Asexual reproduction is the development of a new individual without the

participation of any stages of the sexual cycle, that is, without maturation and
copulation of sex cells with concomitant reduction of the number of chromosomes
in meiosis. As applied to multicellular animals, asexual reproduction means
development of new individuals at the expense of somatic cells-cells of the soma,
or body, as opposed to generative or sex cells (gametes).
The part of the parental organism giving rise to a new individual in asexual
reproduction may be called a blastema, and it always consists of a group of
cells, whereas in sexual reproduction the new individual develops frorm one cell,
the fertilized or parthenogenetically activated ovum. The development that starts
from a blastema may be termed blastogenesis, as opposed to embryogenesis, or
development from the ovum. The individuals resulting from asexual reproduction
are often referred to as blastozooids; individuals developing from an egg are then
termed oozooids.
The blastozooids may have the same general organization as oozooids, or they may
even be indistinguishable from the latter. How this is achieved, in spite of the
profoundly different initial stages, is the second major problem in relation to
asexual reproduction.
Taking the size of the fragment and the degree of its organization as a guiding
principle, we may subdivide the infinite multiplicity of modifications occuring in
different Metazoa into three main types:
1. Fission: The new individual is formed from a relatively large portion of the body
of the matermal organism, and differentiated organs and tissues or their parts are
passed on to the offspring.
2. Budding: The new individual develops from a small outgrowth on the surface of
the parent. No organs of the parent are passed on as such to the offspring, but the
bud is supported by the parental organism at least during the initial stages of its
development.
3. Gemmule formation: The new individual develops from groups of cells which
become completely cut off from the matemal individual and disseminated, so
that the development is, from the start, quite independent of the maternal body.
Fission. The simplest form of fission is the separation of an adult indivdual into
two parts of approximately equal size, similar to the binary fission in protozoans.
In Metazoa it occurs in the most typical form in coelenterates and worms. In some
corals (Anthozoa) fission occurs in the rather rare fom of longitudinal fission, with
the division plane in the long axis of the body. Fission also occurs in some brittle
stars in which parental individuals break across the disc, and each half then
proceeds to regenerate the missing half of the disc and three arms. In the worms
the planes of division are transverse to the body axis, so that the worm is divided
into anterior and posterior halves. This type of reproduction is found in annelids,
both polychaetes and oligochaetes. Both halves inherit from the parent the skin, a
section of the alimentary canal, sections of the nerve cord, and in annelids, a
number of mesodernal segments, its muscles and nephridia, and corespondingly,
sections of parenchymatous mesoden in rhabdocoeles. The posterior individual is
at first devoid of a head and therefore initially lacks the supraesophageal ganglion,
sense organs (eyes), tentacles, and mouth parts, where these occur. The head is
produced either subsequent to division or, more often, in preparation for the
division, so that the posterior individual is fully developed when the two
blastozooids [Link] rhabdocoeles and some oligochaetes, new fissions may be
started by one or both filial individuals even before they are separated from each
other. The result is a chain of blastozooids, some of them in different stages of
reconstitution (degree of head development). The division may also be very
unequal, so that of the twó or several zooids, one may be cleardy offspring.
distinguishable as the parent and the other, or others, as offspring. Cases in which
subsequent transverse divisions occur prior to the earlier divisions being completed
lead to a special form of transverse fission, known as strobilation, in which
numerous transverse divisions occur more or less simultaneously or in close
succession, giving rise to a number of filial blastozooids. This is the classical
method of propagation by which, in Scyphozoa, the asexual polypoid generation
gives rise to the sexual medusoid generation. Similar multiple transverse division
occurs in some polychaetes and also in [Link] the latter, the part undergoing
transverse division is the abdominal section of the body, which is devoid of the
branchial chamber but contains the intestinal loop. Both in worms and in ascidians,
the initial separation of the zooids is performed through the activity of the
ectodemal epidermis. The epidermis forms a circular constricion which cuts
inward, severs the internal organs, and eventually cuts the body of the animal in two
Budding. The most typical examples of budding are found in coelenterates and
tunicates. Superficially, the early bud appears as a small swelling or nodule on the
lateral surface of the body of the parental animal. The nodule grows, takes shape,
and develops a mouth and a whorl of perioral tentacles, in the case of the
coelenterates, or in the tunicates, develops a branchial chamber and other
associated structures including the atrial cavity and the oral and atrial siphons. The
bud may become completely separated from the maternal body or may remain
permanently in connection with the latter, thus leading to the formation of a
colony. In the freshwater hydra, which normally exists in the form of single
polyps, the daughter individuals may remain connected to the matemal body and
occasionally even start forming secondary buds, but eventually this temparary
colony splits into single [Link] always are the buds directly formed on the
body of the parental zooids. Often the buds develop on special outgrowths of the
maternal animals; the outgrouths are then called stolons. These are typicaly present
in many tunicates., but the branches of a hydrozoan colony on which the polyps
develop have the same significance.
Gemmule Formation. This form of asexual reproduction is found in freshwater
sponges and in bryozoans, and in both cases bodies are produced called gemmules
in sponges and statoblasts in bryozoans that can survive after the matermal
individual (or rather matermal colony) dies off during an unfavorable season
(winter in temperate countries, periods of drought in warmer cimales). The
gemmuies and statoblasts are formed in the interior of the parental body from a
number of undifferentiated cells, which become enclosed in a shell with special
spicules in sponges or in a chitinous envelope in bryozoans. Upon destruction of
the parental body, the gemmules or statoblasts are set free, and after conditions
have become favorable, the cells contained inside burst out and develop a new
individual. For the gemmules of sponges,such favorable conditions are created by
the temperature rising above 16C.
COMPARISON OF BLASTOGENESIS AND EMBRYOGENESIS
1. Definition
Blastogenesis: It is the formation of a new individual from a bud (blastema) or cell
mass without the fusion of gametes. It usually occurs in asexual reproduction and
involves direct development from somatic or undifferentiated cells.
Embryogenesis: It is the series of events by which a zygote (fertilized egg)
develops into a fully formed embryo. It involves highly coordinated cell division,
differentiation, and morphogenetic movements.

2. Origin and Initiation

Blastogenesis:- Initiates from buds, blastema, or regenerative tissues.

- Does not require fertilization.
- Example: Budding in Hydra, Regeneration in Planaria, Colonial ascidians.
Embryogenesis:- Initiates after fertilization of gametes (sperm + ovum).
- The zygote is the starting point.

3. Genetic Aspect

Blastogenesis:- Produces offspring genetically identical to the parent (clones).

- No genetic recombination or variation.
Embryogenesis:- Offspring are genetically different from parents due to meiosis
and recombination.
- Promotes genetic diversity.

4. Developmental Stages 5. Examples

Blastogenesis:- Few and simple stages: Blastogenesis:- Hydra → budding
i) Formation of bud/blastema Planaria → regeneration
ii) Cell proliferation and differentiation Colonial Ascidians → blastogenetic
iii) New individual detaches or remains zooids

Embryogenesis:- Complex, multi-step process: Embryogenesis:- Frog, Chick,

1) Cleavage – rapid mitotic divisions of zygote Humans → zygote to embryo

2) Blastulation – formation of blastula development
3) Gastrulation – formation of three germ layers
4) Organogenesis – formation of tissues and organs
5) Morphogenesis – shaping of body plan

6. Complexity 7. Mode of Reproduction

Blastogenesis: Simple, short, direct. Blastogenesis: Asexual reproduction.

Embryogenesis: Complex, long, regulated. Embryogenesis: Sexual reproduction.

8. Occurrence
Blastogenesis: Found in lower invertebrates like Hydra, Sponges, Planaria, colonial
Ascidians.
Embryogenesis: Found in all sexually reproducing animals including invertebrates
and vertebrates.

9. Evolutionary Significance
Blastogenesis:- Ensures rapid multiplication and survival of species.
- Useful in stable environments.

Embryogenesis:- Generates variability and adaptability.

- Allows development of complex body forms.

Sexual reproduction is a biological process in which two haploid gametes

(male and female) fuse to form a diploid zygote, which later develops into a new
individual. It is a universal method of reproduction in higher animals and ensures
genetic variation and evolution.

General Features of Sexual Reproduction

1. Involves the production of haploid gametes by meiosis.
2. Male gamete (sperm) fuses with female gamete (ovum).
3. Fertilization restores diploid chromosome number.
4. Zygote develops into an embryo and finally into an adult.
5. Maintains alternation of haploid (gametes) and diploid (zygote/organism)
phases.

Steps in Sexual Reproduction

1. Pre-fertilization events- Gametogenesis: formation of gametes.

- Gamete transfer: transfer of male gamete to female reproductive tract.

2. Fertilization-Fusion of male and female gametes (syngamy) results in the

formation of a diploid zygote. It may be external (outside the body, e.g., frog,
fish) or internal (inside the body, e.g., reptiles, birds, mammals).

3. Post-fertilization events - Cleavage: zygote undergoes repeated mitotic

divisions.- Blastulation: formation of blastula.- Gastrulation: establishment of
germ layers (ectoderm, mesoderm, endoderm).
- Organogenesis: formation of tissues and organs.

Significance of Sexual Reproduction

- Maintains chromosome number by alternating haploid and diploid phases.
- Introduces genetic variation through recombination and fertilization.
- Enables evolution and adaptability.
- Ensures survival and continuation of species.

Types

GAMETOGENESIS

For successful reproduction the utmost need is the production of functional

gametes by the organism. Gametes are haploid reproductive cells which
carries haploid set (in case of humans n=23) of chromosomes and are
formed from diploid gonads(Testes in case of males and ovaries in case of
females). Gametes are produced by specialized cell division known as
reductional division or meiosis. Although both spermatozoa and egg are
morphologically distinct but both undergo similar stages of development
Gametes are the physical carriers of genetic information which transmit
parental characters from one generation to next generation. In case of
males the gamete is sperm where as in case of females the gamete is
known as [Link] the process of gamete formation is called as
gametogenesis and is of two types:
Spermatogenesis Oogenesis
Spermatogenesis
is the developmental process from germ cell that ends at formation of
haploid spermatid, which begins at the puberty and occurs in the
seminiferous tubules of the testis. Once the process starts, it is a
continuous process and occurs throughout the life of a male. The process
of is divided into three major phases: i. Proliferative phase or Multiplicative
phase, [Link] phase, and iii. Post Meiotic phase (Maturation phase)
Proliferative Phase
It involves the rapid multiplications of primary germ cells which begin with
the arrival of Primordial Germ Cell (PGCs) (also known as Gonocytes) at
genital ridge of the male embryo. These Gonocytes get incorporated into
the sex cord which becomes seminiferous tubules with the course of
development. The Gonocytes gets differentiated into the numerous stem
cells known as type A1 spermatogonia(Spermatogonia are the diploid cells
with chromosome number 46 in case of human beings). These are true
stem cells having the ability to reinitiate the process of spermatogenesis
when transferred to mice whose sperm production is obliterated through
injection of toxic chemicals. Spermatogonia resides in the stem cell niches
at the junction of Sertoli cells (the epithelium of seminiferous tubules), the
interstitial cells (Leydig cells) and the testicular blood vessels. There are
adhesion molecules that join the Spermatogonia directly to the Sertoli cells
which provide nourishment to them throughout the process of development.
This mitotic proliferation of stem cells amplifies the small population of
Gonocytes into a population of type A Spermatogonia which can generate
more than 1000 spermatids per second in adult human male.
Meiotic Phase
The undifferentiated type A Spermatogonia are the sperm stem cells and
that generate another type A Spermatogonia and type A1 spermatogonium
with the onset of puberty. Type A1 spermatogonia have high level of Stra8
transcription factor and are committed to a meiotic pathway. They undergo
five mitotic divisions but keep cytoplasmic connection intact between
themselves. These cells form a syncitum in which each cell communicates
with others via cytoplasmic bridges about 1 micrometer in diameter. There
are chains of 2-32 linked A1 Spermatogonia in human males. These cells
divide to produce typeB Spermatogonia which are the precursors of the
spermatocytes and are the last cells of the line that undergo mitosis. They
divide once to generate Primary spermatocytes that enter into meiosis
The meiotic phase of spermatogenesis is regulated by numerous factors
such as synthesis of BMPs by the Spermatogonia and synthesis of retinoic
acid by the Sertoli cells during puberty. High concenteration of the BMP8b
is needed for the differentiation of the germ cells. Studies have showed that
mice lacks BMP8b factor and thus is not able to induce spermtogenesis at
the age of puberty.
Maturation Phase
In maturation phase primary spermatocytes undergoes the first meiotic
division and produce secondary spermatocytes (chromosome number 23).
During the first meiotic division homologous chromosomes and their non-
sister chromatids undergo the process of crossing over which leads to
introduction of variations in the secondary spermatocyte. Meanwhile, the
secondary oocyte undergo the second meiotic division and produce haploid
cells called as spermatids that remain connected to one another through
their cytoplasmic bridges which get separated from each other during the
process of spermeogenesis. The spermatids are having haploid nuclei but
are functionally diploid. During the division from type A1 spermatogonia to
spermatids the cells move farther and farther away from the basal lamina of
the seminiferous tubules into the lumen of the seminiferous tubules. Thus
each type of cell can be found in a particular layer of the tubule. The
spermatids are located at the border of the lumen where they lose their
cytoplasmic connections and differentiate into spermatozoa. In human the
progression from Spermatogonial stem cells to mature spermatozoa take
65 days The transition between Spermatogonia and spermatocytes is
mediated by the factors release by the Sertoli cells as follows: Glial cell
lines- Derived Neurotrophic Factor (GDNF) Stem Cell Factor (SCF)
GDNF levels determine whether the dividing Spermatogonia last as the
Spermatogonia or enter the meiotic phase to become spermatocytes. Low
level of GDNF is required for the differentiation of Spermatogonia whereas
high level of GDNF is needed for the self renewal of stem cells. SCF
promotes the transition to spermatogenesis. Both GDNF & SCF are
upregulated by FSH, so these two factors serves as link between Sertoli
cells and the endocrine system which provide the mechanism for FSH to
increase the production of sperms.
Post-Testicular Sperm Maturation
Studies have found that ‘major maturation-associated’ sperm membrane
antigen (glycopeptides) appears on the surface of rat spermatozoa during
post-testicular sperm maturation in the distal epididymis of rats. It has been
found through in-situ transcript hybridization,molecular analyses of genomic
DNA fragments and immunohistochemical staining that homologouscounter
parts are present in other mammals, including humans. In case of humans,
this homologue is an abundant epididymal gene product that has been
identified as lymphocyte surface antigen CD52. These Glycosylphosphatid
-yl Inositol (GPI)-anchored glycopeptides (absent in testis) are located
within the epididymal epithelium that are transferred to sperm cell from
epididymal through GPI-anchor; remains intact. Human TRPM 2/ clusterin,
is another gene associated with sperm maturation.
Hormonal Control of Spermatogenesis
Three hormones produced by male gonads directly or indirectly control the
process of spermatogenesis. They are- testosterone, estradiol and inhibin.
Testosterone is secreted by leydig cells which are present adjacent to
seminiferous tubules. Estradiol and inhibin are secreted by sertoli cells or
interstitial cells which are present inside the seminiferous tubules. The
initiation of the process of spermatogenesis greatly depends on the activity
of hypothalamus and adenohypohysis Deficiency of Fibroblast Growth
Factor 2 (FGF-2) leads to abnormal spermatogenesis and altered sperm
physiology. Earlier it was revealed that the presence of Fibroblast Growth
Factor 2 (FGF-2) and its receptors (FGFRs) in human testis and sperms
are involved in spermatogenesis and in motility regulations. Some studies
were carried out to analyze the role of FGF-2 in the maintenance of sperm
physiology using FGF-2 Knock Out (KO) mice experiment which showed
that in Wild-Type (WT) animals, FGF-2 are expressed in germ cells of the
seminiferous epithelium, in epithelial cells of the epididymis and in the
flagellum and acrosomal region of epididymal sperm. This suggest that
FGF-2 exerts a role in mammalian spermatogenesis and the lack of FGF-2
lead to deregulated sperm production as well as altered sperm morphology
and function. FGF-2-deficient mice constitute a model for the study of the
complex mechanisms underlying mammalian spermatogenesis.
 Factors Affecting Spermatogenesis
Temperature: Normal body temperature or above it, sensitize the
seminiferous epithelium. So the optimal temperature (2 degree below body
temperature) is maintained by the scrotum. Scrotal Reflex: Cremasteric
muscles and Dartos smooth muscles in scrotum, position it towards or
away from the heat of the body. Others: Deficiency of vitamin E, B and
A; infectious diseases; X-ray exposure, metals like lead and cadmium,
dioxin, alcohol, pesticides and anabolic steroids, and DNA damage due to
oxidative stress.
SPERM MATURATION
Sperm is the male reproductive cell and is derived from the Greek word
sperma (meaning seed). In the types of sexual reproduction known as
anisogamy and its subtype oogamy, there is a marked difference in the size
of the gametes with the smaller one being termed the ‘male’ or sperm cell.
A uniflagellar sperm cell that is motile is referred to as a spermatozoon,
whereas a non-motile sperm cell is referred to as a spermatium. Sperm
cells cannot divide and have a limited life span, but after fusion with egg
cells during fertilization, a new organism begins developing, starting as a
totipotent zygote. The human sperm cell is haploid, so that its 23
chromosomes can join the 23 chromosomes of the female egg to form a
diploid cell. In mammals, sperm develops in the testicles, is stored in the
epididymis, and released from the penis. The main sperm function is to
reach the ovum and fuse with it to deliver two sub-cellular structures:
~Male pronucleus that contains the genetic material ~ Centrioles that are
structures that help organize the microtubule cytoskeleton
Longer sperm cells are better than their shorter counterparts at displacing
competitors from the female’s seminal [Link] benefit to females is
that only healthy males carry ‘good’ genes that can produce long sperm in
sufficient quantities to outcompete their competitors .
Physiological Maturation of Sperm ~Haploid spermatid is round,
unflagellated cells that undergo transformation [Link] of haploid,
non-motile spermatid into motile elongated spermatozoa.
Spermiogenesis is the process by which haploid round spermatid
completes an extraordinary series of events to become streamlined motile
[Link] begins after spermatocytes complete two
quick successive meiotic reductive divisions to produce haploid round
spermatids. Further no cell division occurs as the spermatids undergo
complex cyto differentiation, over a period of 2–3 weeks in mice and rats to
form mature elongated spermatids that are ultimately released from the
seminiferous epithelium through a process known as spermiation. Main
significance of this process is to increase the sperm motility and to make
sperm more potential so that it could easily penetrate the ovum and
process of fertilization could takes place. During spermiogenesis the
developing sperms keep their heads embedded in the sertoli cells so that
they could easily draw the nourishment from them .There are following
processes involved during changes and maturation of sperms:
Changes in Nucleus: Initially the change that takes place in the process of
spermatogesis is the progressive elongation and reduction in volume of the
nucleus by losing water from the karyolymph. This step is essential, since it
reduces the weight of the spermatozoa and enhances the sperm motility.
Along with this condensation of DNA and reduction of RNA takes place.
Chromatin fibers become closely packed and form uniformly dense
[Link] these changes reduce the volume of nucleus to 0.5 %. In the
compact chromatin, DNA is transcriptionally active therefore protein
synthesis in spermiogenesis utilizes the stored mRNA molecule.
This is an example of Post transcriptional regulation of gene expression.
During all these changes that take place, nucleus attains a characteristic
shape and forms the sperm head. Shape of the nucleus is determined by
the pattern of DNA-protein interaction during the condensation.
Changes in Golgi Complex: Golgi apparatus of spermatid gathers
anterior to the nucleus. Membrane bound pro-acrosomal vesicle develop in
the Golgi apparatus. These proacrosomal granules accumulate into a
single large Acrosomal Vesicle having dense Acrosomal granule in it.
Acrosomal vesicle joins with nucleus and forms the future anterior end of
the sperm. When the acrosomal vesicle attains its size, it assumes the final
shape of mature acrosome.
Changes in Centerioles: Spermatid has two types of nucleus, during the
process of spermiogenesis both the centrioles change their position and lie
behind the nucleus. Out of these two centerioles, one enters into
depression developed in the posterior part of the sperm and latter one is
called as the proximal centriole.
The remaining centeriole is known as distal centriole which will be present
behind the proximal centeriole and its axis coinciding with the longitudinal
axis of [Link] are following functions of centriole:
~ Proximal centeriole enters inside the ovum along with sperm and plays a
role in the spindle formation.~ Distal centeriole give rise to 9+2 axial
filament of the flagellum and act as basal granule.~ In some Mammals
Proximal and Distal Centeriole get disappears after organizing the
connection between neck and middle piece.
Changes in Mitochondria: Mitochondria play an important role in the
process of spermeiogenesis in the middle piece. It gets accumulated
around the proximal part of the axial filament and distal centeriole
Gradually, these mitochondria loses their individuality and fuse together
forming two densely packed bodies, one on the either side of axial filament
(mammals only ) and this sheath is known as Nebenkern (spiral
arrangement of sheath is visible only in mammals ) In other animals,
mitochondria are joined to form massive clumps known mitochondrial
bodies
Changes in Cytoplasm: Most of the cytoplasm in spermatid is lost and
remaining cytoplasm forms a condensed layer in the peripheral region of
the spermatid. This layer is known as Manchette band which surrounds the
middle piece as well as the posterior part of sperm’s head.
Changes in Plasma Membrane: Plasma membrane extends to surround
the acrosome, nucleus and middle piece and tail of the sperm. Initially
some proteins remain embedded over the surface of plasma membrane
and acts as recognition or binding factors during the process of fertilization.
Metamorphic Changes: The different steps of spermiogenesis are
distinguished by the morphological appearance of the developing
acrosome and the change in the shape of the nucleus. During
spermiogenesis, round spermatids having a spherical central nucleus,
begin to assemble the acrosome and the axoneme structures required for
the fertilization and motility respectively. Thus the major event occurring
during spermiogenesis is the assembly of the sperm flagellum. The central
component of the flagellum i.e. microtubule-based axoneme, is assembled
soon after the completion of meiosis. As spermatids elongate, the
accessory structures needed for flagella function (outer dense fibers,
fibrous sheath,mitochondrial sheath) are assembled around the central
axoneme. The final stage of spermiogenesis is known as spermiation and
is the process by which the elongated spermatids undergo their final
remodeling and get released from the seminiferous epithelium. Spermiation
is a complex, multi-step process, which is particularly vulnerable to
disruption. In case of mice, the last stage of sperm maturation is controlled
by Katnal1 gene. This is expressed in sertoli cells which supports and
nurture sperm at the site of spermatogenesis. Dysfunction of Katnal1 gene
leads to male infertility.
Oogenesis
The oogenesis is the process of development and maturation of ovum.
Mammalian oogenesis differs greatly from the spermatogenesis. Eggs
mature through intricate coordination of hormones, paracrine factors and
tissue anatomy. The site of oogenesis is female gonads i.e. ovaries. There
are one pair of ovaries (one on each side) are located in lower abdominal
cavities of female. All the events of oogenesis take place inside ovaries.
Histological, each ovary is made up of a dense fibrous connective tissue
called stroma. It contains blood and lymph vessels,nerves and follicles in
various stages of development. The stroma is surrounded by a layer of
germinal epithelial cells and internal to it is present a thin layer of
connective tissue, called the tunica albuginea. Stroma is further
differentiated into outer broad cortex and inner narrow medulla containing
only blood and lymph [Link] cortex contains ovarian follicles in
various stages of development and regressing follicles like corpora lutea
and corpora albicans and interstitial cells. One follicular cell develops to
become an ovum and the other cells called discus proligerous around it
protect and nourish the ovum. It is attached to one side of the follicle. In a
mature Graffian follicle, a fluid-filled cavity appears which separates an
outer layer of cells, the membrana granulosa, from the cells of discus
proligerous. The follicular cells around the ovum secrete a thick membrane
called zona pellucida. It is covered by another membrane of striated
columnar cells, called corona radiata. Such a mature follicle is known as a
Graffian follicle. The Graffian follicle migrates towards the surface of the
ovary and ruptures to release the ovum (ovulation). The release of ovum
left a mass of follicular cells surrounding a blood clot. Gradually these
follicle cells proliferate rapidly and transform into a yellow colored mass of
cells, the corpus luteum, which produces a hormone,progesterone. It is
responsible for preparatory changes in the uterus for fertilization and
retention of the embryo during [Link], the corpus luteum remains
active throughout the period of embryonic development. If, somehow, no
fertilization occurs, it degenerates leaving a scar,called corpus albicans.
The process of oogenesis is broadly separated into three phases:
Multiplicative phase, Growth phase, and Maturation phase.
Multiplicative Phase or Proliferative Phase
In contrast to spermatogenesis which begins in males at puberty,oogenesis
beginsin females before they are born. During early fetal development, i.e.,
2nd to 7th months of gestation period, Primordial Germ Cell (PGC) migrate
from the yolk sac to developing ovaries. These primordial germ cells get
differentiated within the ovaries to form Oogonia. The oogonia are diploid
(2n) stem cells that divide mitotically to produce roughly 7 million germ
cells. Even before birth most of these germ cells degenerate in a
phenomenon called Atresia.
Growth Phase-Few of oogonia divide mitotically and develop into larger
cells called Primary oocytes. These primary oocytes enter into prophase of
meiosis-I during fetal development. After one meiotic division the oocyte
arrests itself in the Diplotene stage of first meiotic division. This prolonged
arrest in the diplotene stage is also referred as Dictyate resting stage. The
oocytes are maintained in the dictyate resting stage by outer layer of
ovarian follicular cells. During the arrested stage of development each
primary oocyte is surrounded by a single layer of flat follicular cells and
entire structure is called as Promordial follicle. Ovarian cortex surrounding
primordial follicle consists of collagen fibre and fibroblast like stromal cells.
At birth, approximately 200,000 to 20,00000 primary oocytes remain in
each [Link] of which 40,000 are present at puberty whereas around
400 mature and ovulate during women’s reproductive lifetime. Remaining
primary oocytes undergo atresia.
Maturation Phase -With onset of puberty, a group of oocytes periodically
resume meiosis. At that time Luteinizing Hormone (LH) from the pituitary
gland releases the block and permits the oocyte to resume meiotic division.
Primary oocyte completes 1st meiotic division producing 2 haploid cells of
unequal size each with 23 [Link] cell produced by meiosis
are called as first polar body having only nucleus and small amount of
cytoplasm. Larger cell are known as secondary oocyte which receive most
of cytoplasm. Once a secondary oocyte is formed, it begins meiosis-II
which stops at metaphase. Meanwhile, a mature graffian follicles rupture to
release secondary oocyte and the process is known as Ovulation. At
ovulation,secondary oocyte is expelled into pelvic cavity together with first
polar body and corona radiata. Normally, these cells are swept into uterine
tube. If sperms are present in the uterine tube, its penetration takes place
into the secondary oocyte and meiosis II is resumed. Secondary oocyte
splits into two haploid cells of unequal size. Larger cell is ovum or mature
egg and second polar body. If there is fertilization then the nuclei of sperm
cell and ovum unite forming diploid zygote. If fertilization does not occur,
the cells get degenerated. It has been hypothesized that steroids act
directly on the oocyte to induce maturation whereas pituitary hormones act
through the mediation of follicular tissue Evidences in support of
physiological maturation are: A positive response to an activation
stimulus The ability to undergo cleavage and subsequent development
Mechanism of Maturation
Below are the steps that occur in the mechanism of maturation:
Resumption / Reinitiation of Meiosis-I
In amphibian oocyte, the resumption of the meiosis I requires the
[Link] progesterone is secreted by the Follicular cells in
response to the gonadotropic hormone secreted by the anterior lobe of
pituitary. Within 6 hours of progesterone stimulation, the nuclear membrane
dissolves and this is known as Germinal Vesicle Breakdown (GVBD) as the
nucleus in prophase is called as Germinal Vesicle. Along with this, other
events that take place are the retraction of the microvilli, disintegeration of
the nucleoli, contraction of chromosome and the chromosomal migration
towards the animal pole. Promptly first meiotic division and the ovulation
(i.e., release of ovum) take place from the ovaries, when the egg is
released from the ovaries and is in the second meiotic metaphase stage.
Germinal Vesicle Breakdown (GVBD) by Progesterone
M phase of the cell cycle in both (Meiosis and Mitosis) is regulated by
Mitosis Promoting Factor or Maturation Promoting Factor (MPF). MPF has
two units P34 and Cyclin B. P34 protein is a cyclin dependent Kinase and
its activity is dependent on Cyclin. Since all the components of MPF are
present in the amphibian oocyte, it is generally thought that Progesterone
converts inactive MPF into active MPF. Mediator of the progesterone signal
is the C-moss protein. Progesterone reinitiates the meiosis by causing
polyadenylation of maternal C-moss mRNA that has been stored in the
cytoplasm, which is then translated into a 39-KDa phosphoprotein (C-moss
protein). C-moss protein which is only detectable during oocyte maturation
is destroyed quickly upon fertilization. This C-moss protein phosphorylates
the P34 subunit of MPF and also activates the phosphorylation cascade.
Now the activated MPF allows the germinal vesicle breakdown and chromo
-somes to divide. If the translation of C-moss mRNA is inhibited by injecting
C-moss antisense mRNA into ooocyte then there will not be any germinal
vesicle breakdown and hence resumption of meiosis I will not take place.
Resumption of Meiosis-II
After the first meiotic division the oocytes again arrest its division at
metaphase of second meiotic division, this block or arrest is caused by the
CSF (Cytostatic Factor). CSF is a complex of proteins that includes C
moss, Cyclin Dependent Kinase 2(Cdk2) and MAP Kinase, Erp1 (an active
protein which is synthesized immediately after first meiotic division).
Phosphorylation of Erp1 block the degradation of cyclin by Anaphase
Promoting Complex and the arrest at the metaphase stage is broken by
fertilization. If fertilization occurs, calcium ion flux activates Calcium Binding
Protein – Calmodulin which activates two enzymes and these two enzymes
further inactivates CSF, Calmodulin dependent Protein Kinase 2 (Cam
PK2) and Calpain2 (calcium dependent Protease) inactivates C-moss The
maturation is the process that takes the oocyte from the Germinal Vesicle
stage to a stage of meiosis at which the egg can be fertilized or in other
words it’s a process of making the oocyte ready for the fertilization. After
the complete development of oocyte, it enters into second phase where
resumption of meiosis takes place for the maturation of egg. Varying with
different species, the growth of oocyte arrests at different stages of cell
cycle. In ascidians and some Molluscs the growth arrest at Metaphase 1
and in case of Amphibians and Mammals the growth arrests at Metaphase
2 stage of cell cycle. In case of reptiles and birds during the vitellogenic
phase of oogenesis, the oocyte attains its full growth and oocyte become
ready for the next phase of oogenesis, i.e., resumption of meiotic division
which is essential for the final maturation of the oocyte. For the successful
fertilization, the oocyte maturation is pre-requisite event that comprises of
the breakdown of germinal vesicle, chromosomal condensation and
expulsion of first polar body in case of vertebrates. The breakdown of
germinal vesicle leads to the intermixing of nuclear components with the
surrounding cytoplasm. However, in case of animals which do not have
haploid generation, the completion of meiosis is immediately followed by
the fusion of gametic nuclei, i.e., fertilization. In case of reptiles and birds
the first meiosis gets arrested at the diplotene stage of Prophase I of
Meiosis I, where the metabolic activities of oocyte is very high and also
there is enormous increase in the oocyte. In all animals (amphibians,fishes,
reptiles, birds & mammals)the growth of oocyte gets arrested at prophase I
and resumes the meiosis at theend of their growth period.
MATURATION OF EGG
The process in which Prophase I arrested oocyte resume their meiosis is
called as maturation. The term maturation is generally used to describe the
completion of meiosis as two successive meiotic divisions occur in the
animal oocyte, involving the nuclear and cytoplasmic changes. Ovum
maturation or meiotic maturation and the ovulation forms the terminal
stages of oogenesis i.e.,formation of female germ cells. Both nuclear and
cytoplasmic changes occur during oocyte’s final maturation, which have
received relatively much more attention in other animal species than in
reptiles and birds. Ovulation and maturation in vertebrates, including
reptiles and birds are closely linked by common systemic factors which
appear to correlate with the physiology of the ovary and the oocytes or
shows that ovum maturation and ovulation in vertebrates are casually
linked in some manner. It is now well established that administration of
gonadotrophins to female fish, amphibians and mammals having fully
grown oocytes in their ovaries,causes oocyte maturation with concomitant
ovulation In vertebrates, gonadotrophins are secreted by the pituitary gland
or hypophysis. Two different gonadotrophins, such as LH and FSH have
been distinguished in mammals, birds and reptiles except squamates but
only one Gonadotrophic Hormone (GTH) is believed to occur in fish. During
recent years,the effects of gonadtrophic hormones, steroid hormones and
various other chemical substances on oocyte maturation both in-vivo and
in-vitro conditions have been extensively studied in fish, amphibians and
mammals but very little or no work on these lines has been carried out on
reptiles and birds. The results of various in vivo and in vitro studies on
oocyte maturation in fish, amphibians and mammals have suggested that
gonadotrophins act primarily on the follicle granulosa cells to induce
maturation of oocytes in their ovarian follicles .The cell-to-cell interactions
within the vertebrate ovary are of great significance to harmonize the
morphological and functional differentiation of the various cell types (theca
cells, follicle cells and oocyte) in order to ensure the proper development,
differentiation and ovulation of mature eggs which are able to fertilize and
develop normally. Besides producing steroid and non-storoid hormones,
nutrients and possibly the informational molecules for the oocyte, the
follicle cells are also believed to be the source of oocyte maturation
inhibiting factor(s); as suggested for fish and mammals. Actually, a series
of complex interactions between pituitary gonadotrophin(s) and various
steroid hormones secreted by the follicle wall regulate several aspects of
follicular growth and ovum maturation in vertebrates. As a result of the
action of gonadotrophins on follicle cells, the steroid hormone, possibly
progesterone, is believed to be synthesized and released. This steroid
hormone (progesterone) is known to act directly on the oocyte. In response
to progesterone, oocytes undergo a series of morphological and
biochemical changes, resulting in the formation of mature eggs capable of
being fertilized.
Egg Polarity and Symmetry
Literal meaning of polarity is establishment of two opposite poles at both
[Link] is an essential feature of development in all organisms, from
the 1-cell embryo to the establishment of tissue polarity through the whole
organism. It is a characteristic of most cells in metazoan organisms and is
associated with asymmetries in cell shape, protein content, organelle
distribution and ultimately,cell function. Establishment and maintenance of
polarity is of central importance in the process of oogenesis, particularly
during oocyte maturation. Embryonic polarity in Xenopus and Drosophila
are largely dependent upon the localization of mRNA determinants in the
oocyte cortex. Two parts are said to be in symmetry when they appear
identical after a flip, like reflection or mirror. The multicellular organism
has different body parts which are formed from single cell. The distribution
of the interior content of the first cell lay foundation for various
developmental processes which results into formation of different organs
Origin of Polarity
Primary oocyte is an undifferentiated cell in the beginning of growth phase.
When the growth and differentiation of oocyte is taking place inside the
ovaries the polarity develops. Location of oocyte inside the ovary
determines the future animal-vegetal axis of the ovum. Timing of attaining
polarity is different with different species for example in mouse, polarity is
first established in unfertilized oocytes which is characterized by the
formation of metaphase spindle that is localized to the animal hemisphere
of the oocyte. Chromosomal proximity leads to cortex reorganization in a
Ran GTPase-dependent manner and induces actin, myosin 2, Par-3 and
Par-6 accumulation above the spindle. Several studies showed that
different cellular pathways such as Cdc42, Rac-GTPase and the
Mos/MEK/MAP kinase pathway are also involved in the reorganization of
the cortex above the spindle, as they regulate the translocation of the
meiotic spindle to the oocyte cortex which accounts for the polarity of egg
Factors Associated to Polarity of Egg
Uptake of Nutrients: In determining the polarity of eggs, the uptake of
nutrient plays a crucial role. For example in case of Molluscs, the end of
oocyte which is attached to the wall of ovary will become the Vegetal pole
of the egg. Therefore after this observation, it was concluded that the
region of egg which is present in the close proximity of the ovary has more
amount of yolk or in other words more yolk gets concentrated in that region.
For example in case of Mammals and Amphibians they receive the
nutrients from the follicular cells surrounding the ovaries but that doesn’t
turn whole of the egg into the vegetal pole.
Oxygen Consumption: For the establishment of the vegetal pole most of
the researchers believed that the end of oocyte which is present near the
blood vessels gets more oxygen as compared to other end of ovum.
Electrical Field: In 1934 Spek concluded the establishment of the electric
field inside the egg which is responsible for the orientation and movement
ofmolecules entering inside the egg at polar region in accordance to their
electric charge.
Others: There is large number of physical, chemical and environmental
factors that helps in determining the egg polarity. Such as: electric current,
light exposure, surrounding eggs, pH variation, temperature variation, site
of sperm entry, nucleic acids.
Fertilization
is defined as the process of union of
two gametes, eggs and sperm. When mammalian eggs and sperm come into contact in
the female oviduct, a series of steps is set in motion that can lead to fertilization and
ultimately to development of new individuals. Eggs are non-motile, surrounded by
protective egg coverings. These serve to recognize the sperm specifically and prevent
fertilization by more than one sperm (polyspermy). The mammalian egg has zona
pellucida layer around the plasma membrane beneath which cortical granules are
present. The zona pellucida layer makes the egg impenetrable to more than one sperm.
Sperms are highly motile cells consisting of nucleus and mitochondria to provide energy
source and a flagellum for movement. The anterior end of the sperm is highly
specialized which aids in penetration of the egg. Sperms are typically designed to
activate the egg and to deliver their nuclei into the egg cytoplasm via seawater in
marine forms, fresh water in fresh water forms and body fluid in viviparous animals. To
increase the probability of fertilization, the number of sperms must exceed the number
of eggs. Moreover the lifespan of gametes is limited; therefore fertilization must take
place within a short duration of time. Eggs that are shed in water like that of most
invertebrates, fishes and amphibians, have shorter [Link] egg and the sperm are
optimized in opposite ways for the propagation of the genes they carry. The egg is non-
motile and aids the survival of the maternal genes by providing large stocks of raw
materials for growth and development,together with an effective protective wrapping.
The sperm, by contrast, is optimized to propagate the paternal genes by exploiting this
maternal investment; it is usuallyhighly motile and streamlined for speed and efficiency
in the task of [Link] between sperm is fierce and the vast majorities
fail in their [Link] of the billions of sperm released during the reproductive life of a
human male,only a few ever manage to fertilize an egg.
Fertilization and its Types
Fertilization is a phenomenon in which mature male gamete peneteratrate the mature
female gamete ovum and this penetration leads to the fusion of their pronuclei resulting
in the formation of the diploid cellular structure called as Zygote. Fertilization is the
second most important event in the process of sexual reproduction. There are four
stages of fertilization:
Preparation: It includes capacitation and acrosome reaction. The acrosomal
vesicle fusion is the membrane fusion event of this stage.
Binding: It is species-specific interaction of gametes.
 Fusion: Merging of sperm and egg plasma membranes is the membrane fusion
event of this stage.
Activation: It comprises cortical reaction (fusion of cortical vesicles with
the egg plasma membrane) and pronuclear fusion.
The process of fertilization either taking place in surrounding medium outside of body or
in side of body It is a complicated process of sexual reproductioninvolves two gametes:
Eggs are large (~100μm), symmetrical and non-motile cells. Human eggs are arrested
in metaphase of the second meiotic division and complete meiosis only upon
fertilization. Their surface is covered by microvilli. Eggs are surrounded by zona
pellucida which is a glycoprotein coat composed of three glycoproteins(ZPGP I-III). All
three of the glycoproteins contain O- and N-linked oligosaccharides. The zona pellucida
is not an osmotic barrier (in fact, even virus are capable of penetrating it), however it is
a barrier to the sperm. The zona pellucida is the species specific barrier to fertilization
as shown by the hamster experiment. Human sperm are incapable of fertilizing intact
hamster eggs, but can fertilize hamster eggs stripped of their zona pellucida. This is
used clinically to assess the fertilizing capacity of spermSperms: Sperms are small,
asymmetrical and motile cells. They have components like--
Acrosome: is a lysosomal-like compartment derived from the Golgi. It has a low pH and
contains soluble hydrolases (serine protease acrosin).In cross-section through the head
of a sperm, one would cross four membranes in traversing from the plasma membranes
to the nuclear membrane. During the acrosome reaction, fusion of the outer acrosomal
membrane with the plasma membrane releases the contents of the acrosome and
exposes inner acrosomal membrane as functional outer boundary of the sperm head.
Head: It contains the spermatic haploid nucleus. Overlaying the head is a membrane
bound vesicle, the acrosome. Sperm do not possess any organelles associated with
protein synthesis (Golgi body, RER or lysosomes). The sperm plasma membrane is
also highly differentiated and contains proteins localized in distinct regions. One of
these, termed PH-30 or fertilin, is localized in the equatorial region of the sperm and is
involved in sperm-egg plasma membrane fusion.
Middle Piece: At the proximal portion of the tail. Mid piece contains a sheath of
mitochondria, which produce the ATP necessary for the beating of the tail.
Tail: Also referred to as the principal piece. The tail contains the flagellar apparatus,
which is composed of ‘9 + 2’ microtubules and accessory structures. The sliding of the
microtubule is powered by the protein dynein.
Types of Fertilization
The fertilization process in animals can occur either internally or externally, a difference
which is largely determined by the method of birth.
Internal Fertilization: Animals which use viviparous and ovoviviparous reproduction
(embryos develop within the animal’s body) and oviparous animals which lay hard
shelled eggs, use internal fertilization. Internal fertilization involves the union of sperm
and eggs within the body of the (usually female) parent. For internal fertilization to
occur, the male must implant his sperm into the female reproductive tracts. Implantation
can be achieved by either: Copulation, in which sperm transfer is performed by insertion
of the penis or other male intromittent organ and ejaculation into the vagina, or cloaca,:
or by a cloacal kiss, in which two birds press their cloacae together and sperm transfer
takes place. Some animals, such as molluscs, arachnids, salamanders and certain
insects, transfer a spermatophore, a bundle or capsule containing sperm, which is
stored within the cloaca until oviposition takes place.
External Fertilization: Animals which are oviparous produce eggs which are lacking, or
have thin egg membranes and reproduce by external fertilization. External fertilization is
a reproductive strategy involving the joining of gametes outside of the body, either in a
spawning event, where gametes from both sexes are rapidly released into an aquatic
environment, or may occur when eggs are laid by a female on a substrate and are
subsequently fertilized by a male. External fertilization holds certain benefits, such as
reducing chance of contracting sexually transmitted diseases, protection from violent
behavior between organisms and increasing the genetic variation within a population.
Mechanism of Fertilization
The fertilization is a complicated four steps process as discussed below
Step-I Preparation of Sperm
Ejaculated sperms are not ready to fertilize an egg when they enter the vagina. In
response to the dilution of semen in the vagina, they undergo several changes,which
are collectively known as capacitation that includes:
Intracellular Ca++ levels increases. Spermatic motility is activated and tail changes
the beating frequency. Sperm cell surface antigens are lost. The loss of these
proteins renders the sperm more receptive for binding to the egg.
Step-II Sperm-Egg Binding
The process of sperm-egg binding was first studied and understood in invertebrates.
In sea urchins, the sperm head binds directly to the egg’s outer surface and this
triggers the acrosome reaction. The acrosomal contents are released and there is a
balanced Na+ influx and H+ efflux, causing an increase in pH. The increased pH
triggers the dissociation of the profilactin complex (actin and profilin) and the released
actin monomers polymerize to form a filament called the acrosomal process. This
acrosomal process penetrates the egg coatings to allow fusion of the sperm and egg
plasma membranes. In sea urchins then, the sperm literally skewers the egg. In humans
the process of sperm-egg binding is not so [Link] complicating factor is the thick
zona pellucida, which keeps sperm from bindingclose to the egg membrane.
Many studies support a role of galactosyl transferase as a sperm protein involved
in sperm-egg binding; however, other proteins may also be involved. A recent
genetic knockout of galactosyl transferase in mice yielded mice that were completely
fertile and showed normal sperm-egg binding.
Acrosome Reaction: As a result of irreversible binding of the sperm to the egg, the
zona pellucida triggers the acrosome reaction. The outer plasma membrane of the
acrosome fuses at multiple sites with the plasma membrane and the contents of the
acrosome are released. Two of the important components are acrosin, a serine
protease and N-acetylglucoaminidase. Acrosin bores a hole in the zona pellucida so
that the sperm can reach the egg itself. N acetylglucoaminidase hydrolyzes the O-linked
oligosaccharides in ZPGP III to allow the sperm to [Link] a result of the membrane
fusion, a new surface is exposed on the sperm (the inner acrosomal membrane) and
this is thought to contain new binding sites for ZPGP II.
Step-III Sperm-Egg Fusion
For many years the process by which the plasma membrane of the sperm and egg
fused, was a black [Link] and sequencing of Fertilin revealed that á subunit had a
hydrophobic domain that resembled with those on viral proteins that are known to be
involved in membrane fusion. The â-subunit had a disintegrin domain. Disintegrins were
first discovered in snake venom and act as competing ligands for integrins (for example,
snake venom disintegrins, block platelet aggregation mediated by integrins). Fertilin was
one of the first proteins of a family known as ADAMs proteins (for A Disintegrin and
Metalloprotease containing protein) that are involved in cell-cell recognition and cell
fusion events. Although the mechanism for how fertilin causes sperm-egg membrane
fusion is not known, studies have supported its role in membrane fusion. For example, a
peptide corresponding to the viral fusion peptide of á-fertilin is capable of fusing model
membrane vesicles and the disintegrin domain of â-fertilin will block sperm-egg fusion.
The egg integrin involved in sperm-egg fusion (the receptor for the â-subunit disintegrin)
is known to be á6-â1. Once the sperm fuse with egg, the beating of tail stops
immediately. The sperm instead is drawn into the egg by elongation and fusion of the
egg’s [Link] a result, the sperm nucleus and other organelles are incorporated
into the egg cytoplasm. The sperm nucleus undergoes a series of changes, including
chromatin decondensation and formation of a new nuclear envelope, to form a male
[Link] male pronucleus uses microtubules to migrate to the center of the cell,
where it fuses with the female pronucleus to reconstitute a diploid nucleus. Other sperm
organelles (for example, mitochondria) persist during early cleavage stages of the
embryo and it is conjectured that they may play a role in development.
Step- IV Activation Response of Egg
The immediate events after fertilization include the egg’s effort to prevent polyspermy.
Polyspermy refers to the fertilization of the egg by more than one sperm, resulting in
zygotes with greater than a diploid amount of DNA. This causes early embryonic
defects and arrest of development. After sperm-egg fusion, the egg mounts the cortical
reaction to prevent polyspermy. In all eggs, residing just under the plasma membrane
there are membrane bound vesicles known as cortical granules. When a single sperm
penetrates the egg, the cortical granules adjacent to the site are triggered to fuse with
the plasma membrane, exocytosing their contents into the perivitelline space (the space
between the plasma membrane and the zona pellucida).The cortical reaction is
propagated over the surface of the egg by a wave of Ca ++.This was shown by the
aequorin experiment in which the photoprotein aequorin phosphoresced in a wave from
the site of sperm penetration of the egg. As a result of the cortical reaction, two
important enzymes are released into the perivitellinespace:
Ovoperoxidase: In sea urchins, ovoperoxidase catalyzes the crosslinking of tyrosine
residues in the extracellular matrix. This makes the extracellular matrix tough and
insoluble (analogous to the tanning of leather) and a physical barrier is formed which
prevents other sperm from fertilizing the egg. In mammals, ovoperoxidase does not
catalyze tyrosine cross-linking to the point of insolubility. In mammals, its major effect is
thought to be as a spermicial agent.
Hydrolase: Remember Wassarman’s result showing that zona pellucid from fertilized
eggs was incapable of blocking fertilizationfrom fertilized eggs was incapable of
blocking fertilization? Another cortical granule that is released is a specific hydrolase,
which degrades O-linked oligosaccharides on ZPGP III. This renders the Zona Pellucida
incapable of binding additional sperm, thus preventing polyspermy. Activation of the egg
also includes the initiation of development of the new [Link] synthesis and
other metabolic processes are upregulated to provide for thedeveloping embryo.
Species Specific Fertilization by Fertilizin-Antifertilizin Reaction
One of the characteristic feature of fertilization is species specificity, i.e., spermatozoa
of one species will only fertilize the ovum of that particular species only. Species
specificity is of greatly biological importance as it helpful in maintaining the individuality
of that species. This is achieved due to presence of specific chemical substances that
are present on the surface of both the [Link] Fertilizin secreted by the ovum and
Anti-fertilizin secreted by sperm for specific interaction which is essential for fertilization.
Fertilizin is a sperm –agglutinating agent produced by an ovum and plays an important
role in the preliminaries of fertilization. Chemically it is glycoproteinaceous in nature
composed of carbohydrates (Glucose+ Fructose+ Galactose) and amino acids. Specific
interaction of fertilizin takes place with other chemical substance secreted by sperm
known as Anti-fertilizin that causes the sperm to adhere to egg and penetrate. Anti-
fertilizin is composed of acidic amino acids such as Glutamic acid and Aspartic acid.
The interaction between both these chemical substances makes the spermatozoa stick
to egg surface therefore this adhesion of spermatozoa to egg of the same species
through a chemical recognition is called as Agglutination [Link] the
ejaculation inside female genital tract mammalian sperm undergo a series of
biochemical and physiological changes and are called as capacitation and these
changes takes place in the female reproductive tract prior to the Acrosome Reaction
(AR). Intracellular calcium plays an important role in bringing these changes in sperm.
Ejaculated mammalian spermatozoa reside inside the female genital tract for several
hours before acquiring the ability to fertilize the egg. In humans sperm must move out of
the seminal plasma immediately after ejaculation and appear in the fallopian tube within
minutes. These changes involve molecules absorbing on,or integrating into, the sperm
plasma membrane during epididymal [Link] removal or alteration of these
molecules prepares the sperm toward successful binding to the egg and fertilization.
During mammalian fertilization, the capacitated spermatozoon penetrates the cumulus
oophrous of the ovum, and then binds to the Zona Pellucida (ZP) with its plasma
membrane intact. After binding to the egg ZP, the spermatozoon undergoes an
exocytotic process called the Acrosome Reaction (AR). This event is required for
fertilization, because it enables passage of the spermatozoon through the ZP and its
subsequent fusion with the egg [Link] capacitation includes multiple physiological
and biochemical modifications. The biochemical changes associated with the
capacitation process include:
. An efflux of cholesterol from the plasma membrane leading to an increase in
membrane fluidity permeability gets increased to bicarbonate and calcium ions
Hyperpolarization of the plasma membrane
Changes in protein phosphorylation and protein kinase activity
Increase in bicarbonate (HCO3–) concentration and intracellular pH
Ca2+ and cyclic Adenosine Monophosphate (cAMP) levels also gets increased
Generally the process lasts for 6 hours inside the female genital tract.
Activation of Egg and Egg Metabolism
In Sea urchin the activation of egg and egg metabolism involves two mechanisms:
Early Responses: The activation of all eggs depends on an increase in the
concentration of free calcium ions within the egg. Such an increase can occur in two
ways either calcium ions can enter the egg from outside, or calcium ions can be
released from the endoplasmic reticulum within the egg. Both processes vary in
different species. In snails and worms, large amount of calcium probably enters the egg
from outside, while in fishes, frogs, sea urchins, and mammals, most of the calcium ions
probably come from the endoplasmic reticulum; however in both cases, a wave of
calcium ions sweeps across the egg, beginning at the site of sperm-egg fusion.
Presence of calcium ions is essential for activating the development of the embryo.
Studies showed that the calcium-chelating chemical EGTA is injected into the sea
urchin egg; there is no cortical granule reaction, no change in membrane resting
potential and no reinitiation of cell division. Conversely,eggs can also get activated
artificially in the absence of sperm by procedures that release free calcium into oocyte.
Steinhardt and Epel in (1974) found that injection of micromolar amounts of the calcium
ionophore A23187 into a sea urchin egg elicits most of the responses characteristic of a
normally fertilized [Link] elevation of the fertilization envelope,a rise of intracellular
pH, a burst of oxygen utilization and increases in protein and DNA synthesis are all
generated in their proper order. In most of these cases,development ceases before the
first mitosis because the egg is still haploid and lacks the sperm centriole needed for the
division. Calcium release activates a series of metabolic reactions. One of these is the
activation of the enzyme NAD+ kinase, which converts NAD+ to NADP+. This change
may have important consequences for lipid metabolism, since NADP+ (but not NAD+)
can be used as a coenzyme for lipid [Link], the conversion of NAD + to
NADP+ may be important in the construction of the many new cell membranes required
during cleavage. Another effect of calcium release involves oxygen consumption. A
burst of oxygen reduction(to hydrogen peroxide) is seen during fertilization and much of
this ‘respiratory burst’ is used to crosslink the fertilization envelope. The enzyme
responsible for this reduction of oxygen is also NADPH-dependent. Lastly, NADPH
helps regenerate glutathione and ovothiols, which may be crucial for scavenging free
radicals that could otherwise damage the DNA of the egg and early embryo.
Late Responses: Immediately after increase in the levels of calcium ion in sea urchin
egg, its intracellular pH also increases and the rise in intracellular pH begins with a
second influx of sodium ions, which causes a 1:1 exchange between sodium ions from
the seawater and hydrogen ions from the egg. This loss of hydrogen ions is responsible
for the rise in the intracellular pH. It is thought that the pH increase and the calcium ion
elevation act together to stimulate new protein synthesis and DNA synthesis. If one
experimentally elevates the pH of an unfertilized egg to a level similar to that of a
fertilized egg, DNA synthesis and nuclear envelope breakdown ensue just as if the eggs
were fertilized. The late responses of fertilization brought about by these ionic changes
include the activation of DNA synthesis and protein synthesis. In sea urchins, a burst of
protein synthesis usually occurs within several minutes after sperm entry. This protein
synthesis does not depend on the synthesis of new messenger RNA; rather, it utilizes
mRNAs already present in the oocyte cytoplasm. These messages include mRNAs
encoding proteins such as histones, tubulins, actins, and morphogenetic factors that are
utilized during early development. Such a burst of protein synthesis can be induced by
artificially raising the pH of the cytoplasm using ammonium ions.
Cleavage
Process of cleavage was first observed by Swammerdam in 1738 in the egg of frog.
First two cleavage planes in case of toad’s egg were described by Spallanzani in 1780.
The entire process of cleavage in frog’s egg was studied by Prevost and Dumas in
1824. Cleavage is actually the result of two coordinated processes. The first of these
cyclic processes is karyokinesis, the mitotic division of the [Link] mechanical
agent of this division is the mitotic spindle, with its microtubules composed of tubulin
(the same type of protein that makes up the sperm flagellum).The second process is
cytokinesis, the division of the cytoplasm. The mechanical agent of cytokinesis is a
contractile ring of microfilaments made of actin (the same type of protein that extends
the egg microvilli and the sperm acrosomal process).The mitotic spindle and contractile
ring are perpendicular to each other and the spindle is internal to the contractile ring.
The contractile ring creates a cleavage furrow, which eventually bisects the plane of
mitosis, thereby creating two genetically equivalent [Link] actin
microfilaments are found in the cortex of the egg rather than in the central cytoplasm.
Under the electron microscope, the ring of microfilaments can be seen forming a distinct
cortical band 0.1 m wide. This contractile ring exists only during cleavage and extends
8–10 m into the center of the egg. It is responsible for exerting force that splits zygote
into blastomeres; if it is disrupted, cytokinesis stops. Schroeder (1973) has proposed a
model of cleavage wherein the contractile ring splits the egg like an ‘intercellular purse-
string,’ tightening about the egg as cleavage continues. This tightening of the
microfilamentous ring creates the cleavage furrow. Microtubules are also seen near the
cleavage furrow(in addition to their role in creating the mitotic spindles), since they are
needed to bring membrane material to site of membrane addition. Although
karyokinesis and cytokinesis are usually coordinated, they are sometimes separated by
natural or experimental conditions. In insect eggs, karyokinesis occurs several times
before cytokinesis takes place. Another way to produce this state is to treat embryos
with drug cytochalasin B. This drug inhibits the formation and organization of microfila
ments in contractile ring, thereby stopping cleavage without stopping karyokinesis
Planes of Cleavage
During early cleavage, distinct geometrical relationships exist between the blastomeres
i.e., each plane of cell-division bears a definite relationship with each other. There are
different plains of cleavage based on the types and groups of eggs:
Meridional Plane of Cleavage: When a furrow bisects both the poles of the egg passing
through the median axis or centre of egg it is called meridional plane of cleavage. The
median axis runs between the centre of animal pole and vegetal pole.
Vertical Plane of Cleavage: When a furrow passes in any direction (does not pass
through the median axis) from the animal pole towards the opposite pole.
Equatorial Plane of Cleavage: This type of cleavage plane divides the egg halfway
between animal and vegetal poles and line of division runs at right angle to median axis.
Latitudinal Plane of Cleavage: This is almost similar to the equatorial plane of cleavage,
but the furrow runs through the cytoplasm on either side of the equatorial plane.
Types of Cleavage
Considerable amount of reorganization occurs during the period of cleavage and the
types of cleavage depend largely upon the cytoplasmic contents. Different types of
cleavage encountered in different eggs are catalogued below
On the Basis of Amount of Distribution of Yolk
Yolk is a heterogenous chemical substance that retards the cleavage furrows which
divide the cytoplasm. An enormous amount of yolk tends to displace the mitotic
apparatus to an off centre position hence the amount and distribution of yolk greatly
determines the different types of cleavage in different organisms.
i. Holoblastic or Total Cleavage: In this type of cleavage the zygote and blastomeres
are divided completely by cleavage furrow or in other words when the cleavage furrows
divide the entire egg. This kind of cleavage occurs in different types of ovum such as
alecithal, homolecithal and little [Link] this kind of cleavage is of two types:
equal holoblastic cleavage and unequal holoblastic cleavage, discussed below:
Equal Holoblastic Cleavage : When the cleavage furrow cuts the egg into two
equal cells and result into the production of the nearly equal –sized blastomeres
throughout the whole cleavage process because the mitotic apparatus is formed near
the centre of the zygote or the blastomere. It may be radially, bilaterally, spirally or
irregular in symmetry. Found in the alecithal egg of rabbit and microlecithal egg of sea
squirt.
Unequal Holoblastic Cleavage: This kind of cleavage results into the formation of
two different types of blastomeres of unequal [Link] is small sized blastomeres called
as micromeres at the animal pole and large sized blastomeres called as macromeres at
the vegetal pole. Found in telolecithal eggs of frog and bony fishes. In these eggs,the
yolk displaces the mitotic apparatus into animal hemisphere and cleavage furrow is
more prominent in the animal hemisphere as compared to vegetal hemisphere that’s
why numerous and smaller micromeres are found in this region.
ii. Meroblastic Cleavage: In this type of cleavage, mitotic divisions occur only in the
metabolically active cytoplasm which remains confined to the animal or the peripheral
region of the egg. However the yolk remains undivided and this kind of cleavage
generally found in macrolecithal eggs or in other words, when segmentation takes place
only in a small portion of the egg resulting in the formation of blastoderm, it is called
meroblastic [Link] the blastoderm is present in the animal pole and the
vegetal pole becomes laden with yolk which remains in anticleaved state, i.e., the plane
of division does not reach the periphery of blastoderm or blastodisc. Furthe rmeroblastic
cleavage is of two different types: discoidal cleavage and superficial cleavage:
Discoidal Cleavage: This type of cleavage is the characteristic feature of
(meiolecithal eggs) reptiles, birds and other egg laying mammals. A rapid mitotic
division occurs only in the cytoplasmic disc present in the animal pole of the egg
resulting in the formation of different layer of cells. Also, in such eggs there is
displacement of mitotic apparatus towards the cytoplasmic disc under influence of yolk.
Superficial Cleavage: This type of cleavage is the characteristic feature of
centrolecithal eggs of insects. Nucleus also known as the synkaryon is present in the
central mass of the cytoplasm, called as ‘energid’ at the middle of the yolk mass and
undergo many repeated mitotic divisions to form large number of daughter nuclei and all
of these daughter nuclei migrates through the cytoplasmic connections towards the yolk
free cytoplasm present at the peripheral region of the egg. This whole process makes
an egg a syncytium. Later on the mitotic divisions takes place and form many
multinucleated blastomeres arranged around the yolk. However only few nuclei remain
inside central cytoplasm for the digestion of the yolk. Relatively thick region of periplasm
present at the posterior region of the pole called as the Pole Plasm get cleaved into the
pole cells after receiving some energies On the basis of Orientation of Mitotic Apparatus
There are various types of cleavage reported in different group of animals on the
basis of mitotic orientation:
Radial Cleavage: In this kind of cleavage mitotic apparatus is either parallel or
perpendicular to the animal-vegetal axis of the egg and cleavage furrow appears in a
manner that it lies radial to the primary axis of the egg. Radial cleavage mostly
observed in 8- celled stage where the upper blastomeres lie directly above the lower
four cells. Any line passing through the animal vegetal axis of the egg divides the egg
into symmetrical halves. It is greatly observed in the sponges, coelenterates and
echinoderms such as starfish.
Spiral Cleavage: In this cleavage the mitotic apparatus is inclined with respect to
animal-vegetal axis of the egg and the blastomeres arranged in the spiral manner
around the axis. When the mitotic apparatus is inclined /tilted in clockwise direction then
after third cleavage the upper micromeres is displaced obliquely to the right side of
lower tier of micromere. This right handed or clockwise displacement of micromeres is
called dextral spiral cleavage. In some species, the mitotic apparatus is inclined towards
anticlockwise direction than the micromeres are displaced obliquely to the left side of
the macromeres. This left-handed or left side inclination of the macromeres is called as
the sinistral spiral cleavage.
Bilateral Cleavage: In this type of cleavage the mitotic apparatus is present either
perpendicular or parallel to the animal-vegetal axis of the egg and the resulting
blastomeres lies in the radial manner about the primary egg axis as in case of radial
cleavage. But in this case the right and left side of the embryo are distinguishable and
only one plane can divide it into the similar halves. It occurs in case of molluscs, fishes,
amphibians etc.
Rotational Cleavage: In this kind of cleavage first two blastomeres rotates about at
an angle of 90 degree with respect to each other before the commencement of second
cleavage which occurs in two planes, i.e.,meridional in one blastomere and equatorial in
another. It occurs in the wide range of nematodes.
Types of Cleavage on the Basis of Potentiality of Blastomeres
Determinate Cleavage: In Animals showing spiral cleavage, their resultant
blastomere’s fate is fixed right from the beginning of first division of the zygote.
Consequenlty complete embryo will be formed only if all the blastomeres remain
adhered together; if any of the blastomeres looses the contact with another then their
fate deciding ability also gets lost.
Indeterminate Cleavage: During this kind of cleavage the fate of blastomeres is not
fixed; in other words they can form any part of [Link] this kind of cleavage is
found in Ascidians. Effects of Yolk in [Link] fertilized egg in most cases contains
yolk, which are inert bodies. During division these bodies exert mechanical influences.
In the egg of Amphioxus, the yolk is thin and remains uniformly distributed. Therefore
the division is complete and early divisions occur at a very quicker rate. The amphibian
egg contains yolk which is localized at the vegetal pole. Here division gets initiated from
the animal pole and extends towards the vegetal pole, where the progress of cleavage
slows down considerably. Consequently, the animal pole divides faster than the vegetal
pole. The eggs of reptiles and birds are fully laden with large masses of yolk, thus
restricting the cytoplasm and nucleus on the periphery as a circular disc on the animal
pole. The lines of cleavage divide only the small animal pole region. Such effects of yolk
on cleavage pattern influence the pattern of further development.
Mechanism of Cleavage
The incidence of cleavage provides unique opportunity to study the mechanism of cell
division and specially the role of different cell organelles during [Link] differ
regarding the accumulation of force for the initiation of cleavage and following factors
are believed to be responsible for controlling the cleavages:
Localized expansion of cortex Increased stiffness of the cortical cytoplasm
Increase of tangential force activity in the cortex Contractile nature of the regions
near the cortex Formation of new cell membrane from the sub cortical cytoplasm.
Though the above mentioned factors are not clearly understood, it is evident that three
structures present within the cell: Cortical layer, Spindle structures and Chromosomes
play the important part. The energy which is required during the process is supplied by
the metabolic activity of the developing egg. Besides the factors involved in
segmentation, there are cleavage laws which govern the behaviour of the cells during
cleavage. These are: Sach’s rules (The blastomeres tend to divide into identical
daughter cells and a cleavage furrow tends to cut the previous cell at right angles);
Hertwig’s laws (The position of nucleus is vital and it tends to lie at the centre of the
protoplasmic content of the cell. The nucleus exerts influence on cleavage. The long
axis of mitotic spindle usually coincides with the long axis of the protoplasmic content.
During cleavage the long axis of the protoplasm has the tendency to cut transversely);
and Balfour’s law (The rate of cleavage is inversely proportional to the amount of yolk
material present in the egg).Chemical Changes During Cleavage Significant chemical
changes go on in the fertilized egg during cleavage. These changes are:
Increase of Nuclear Material: During cleavage a steady increase in nuclear material
(predominantly DNA) is observed. Cytoplasm of the egg is the source of such nuclear
material. Cytoplasmic DNA contained in mitochondria and yolk platelets are available.
RNA Synthesis: During cleavage messenger RNA (mRNA) and transfer RNA (tRNA)
are synthesized during cleavage, especially in late stages. Synthesis of Proteins:
Throughout periodofcleavage there is steady&spectacular increase in protein synthesis.
Regeneration
You may have observed that when a predator grabs a lizard by its tail, the latter
evades capture by simply leaving the distal part of its tail in the grip of the former.
The tail which moves for a while, apart from being unnerving for the predator also
allows the lizard to escape. The tailless lizard is not unduly worried about the loss
of parts of its tail, as it has the ability to develop the lost part,though imperfectly,
by the mechanism of [Link] is a fascinating phenomenon. It
involves continuity of the developmental processes or reawaking of the process of
morphogenesis and differentiation in post-embryonic life in an already formed and
functional organism. Regeneration occurs at various level of organization. At the
sub-cellular and molecular level it is manifested in the continuous synthesis to
replenish used up substances in the cells. At the sub-cellular and tissue levels it
involves replacement of worn out cells, repair of damaged tissues and healing of
[Link] these levels the ability to regenerate is a universal characteristic of all
animals without which life of any individual would be impossible. At the
organismic level regeneration consists of de novo (afresh) development to restore
the lost part of an organ or the reconstitution of the whole body from the residual
pan of the organ concerned. This involves retracing many of the complex steps of
the original ontogenetic development in a functional body under quite different
physiological and environmental conditions. The capacity for this type of
regeneration is referred to as reparative or restitutive regeneration and is unevenly
distributed in the animal kingdom. Some have great powers to restore lost parts, or
even to form a whole body from a small piece. Others have variously restricted and
limited abilities of such regenegation, and still others have no power of reparative
regeneration at [Link] reasons for such inequality of regenerative power among
animals are not clear. In many groups, the animals exhibit the phenomenon of
autotomy, by which they themselves cast off or lose one or more parts of the body
when disturbed or threatened by an enemy or a predator. The autotomized (self-
amputated) parts are subsequently regenerated. Different animals employ diverse
method for the regeneration of lost parts. The study and investigation of the
phenomena of regeneration are of great help in the efforts to understand the basic
processes and mechanisms of development as [Link] ability has been
examined in a large number of animals belonging to almost every phylum since
this phenomenon was discovered more than 250 hundred years ago. The most
favourite animals for analytical studies to understand the variousdevelopmental
aspects of regeneration have been the amphibians (limb, tail, eye and lens
regeneration) among vertebrates and hydra (coelentrata) and planarians (flat
worms) among the invertebrates. Annelids, arthropods and some others also have
received a fair amount of attention.
TYPES OF REGENERATION
The three basic types of regeneration that occur in animal are:(i) Physiological
regeneration. (ii) Reparative regeneration. (iii) Compensatory hypertrophy.
Physiological regeneration>This type of regeneration is a regular physiological
function involving the continuous replacements of cells and tissues, and so is
indispensible for the maintenance of life in all animals. It is a primary attribute of
all living systems. Without such regeneration there would be no life, as the very
maintenance of an organism depends upon the incessant turnover by which all
tissues and organs renew themselves. For example,regular replacement of
epidermal skin layers and RBC etc., in our body is a must. In some instances quite
substantial quantities of tissues are replaced periodically,as in the successive
production of follicles in the ovary or the moulting and the replacement of feathers
and hairs. In our body one per cent of the total 25Xl0l2 RBCs present in active
circulation, die everyday and so are replaced [Link] time span of regeneration
varies; as for example, mammalian skin epidermal cells,produced at the basal level
may take several weeks to reach the outer surface and be sloughed off, while the
life span of an individual epithilial cell in the intestine may be limited to a few
days. The motile hairlike flagella and cilia of single-celled organisms have the
ability to regenerate within an hour or two after amputation.
Reparative regeneration>This kind of regeneration, as the name suggests,
involves repair of a wound or replacement of a body part removed intentionally or
due to [Link] type of regeneration may include restoration of parts of an organ
or an organ as in regeneration of eye and lens in amphibians or parts of the whole
organism as in limbs of urodeles, or it may be the regeneration of an entire
organism from a pan detached from the parent body as you will see in hydra. The
power of this type of regeneration is not found uniformly in all animals. Some have
great powers of such regeneration;in others it is limited to varying degrees and in
yet others it is not found at [Link] unit deals primarily with the phenomenon of
reparative regeneration, generally referred to simply as regeneration as found in
various invertebrates and vertebrates.
Compensatory hypertrophy>It has been observed that the exact replacement of a
part or organ or tissue is not the only way to regenerate in animals;many of the
body's internal organs compensate for their loss by enlarging what remains, instead
of regrowing the missing [Link] process, called compensatory hypertrophy, is
possible as the remaining mass is usually as good as that which was lost. Liver
regeneration in mammals is a well documented example of this process, where the
size of the residual lobes expand,thus restoring the original mass of hepatic tissues
as well as its functionCompensatory hypertrophy in liver is accompanied by
hyperplasia of its cells and of the histological functional units into which they are
organized. Similar mechanisms have been noted in many endocrine and exocrine
glands following surgery or physiological [Link],thyroid,adrenals
and ovaries are the other organs which regenerate by compensatory hypertrophy.
The way they compensate for such loss is the same way they grow during
[Link], not all organs are able to multiply their functional units in such
a [Link], muscles, lungs and kidney are unable to do so.
PATTERNS OF REPARATIVE REGENERATION
Two different patterns or modes of regeneration can be distinguished. These two
patterns were described by Morgan in 1901 as: a) Epimorphic Regeneration-
Epimorphosis b) Morphallactic Regeneration or morphallaxis.
Epimorphic regeneration:In this type of regeneration the lost part is reformed
and restored by the growth of a bud or blastema from the remaining part of tlie
organism followed by the blastema's redifferentiation and morphogcncsis.A much
studicd exalnple is salamander limb [Link] brief, in this type of
regeneration,a bud or blastema is formed at the site of amputation. The blastema
contains the cells that will form the regenerated part, and is encountered in the
rcgencrative processes of all animals in which epimorphic regeneration occurs. The
blastema is made up of cells that look very much alike despite their diverse origins
froni different tissues of the residual stump
Morphallaxis Regeneration:This type of regeneration occurs in plants, sponges
and coelenterates such as jelly fishes and hydra. The missing parts are replaced by
reorganization or remodelling of the pre-existing ones. The wound is healed and
the neighbouring tissues reorganize themselves into whatever parts may have been
lost or removed. Thus in this type of regeneration the residual part of the animal is
capable of restoring the lost part or giving rise to the entire organism just by
remodelling or reorganising the entire available mass of cells into a new [Link]
process does not involve growth until the lost part or the whole body is
regenerated, which is necessarily small at [Link] to attain normal size occurs
later. Morphallactic regeneration can occur in complete absence of cell division, as
is seen in the case of regeneration in Hydra.A part of Hydra as small as lJ200th of
the original individual can form a complete animal without cell proliferation being
involved. Thus even a few cells are capable of forming a new organism; Similarly
in sponges a few archaeocytes are capable of regenerating a whole sponge body.
The morphallactic process of regeneration is observed only in lower groups of
animals. Animals with more complex organization regenerate their parts
differently,usually by the production of a specialized bud or blastema.
Compensatory regeneration:This type of regeneration occurs when some of the
part of any organ damage and it regenerates by the proliferation of existing tissue.
In this the cells divide, but do not undergo [Link] cells are
produced but do not form a mass of undifferentiated [Link]: Regeneration
in the mammalian liver.
Autotomy:It is the process of detachment of the body part when being threatened
by [Link] lost part then [Link] process is also called selfmutilation.
Example:Crabs break off their leg on approaching of the enemy Holothurians
throw off their internal viscera,Starfish breaks off an arm,Lizards detach their tail.
Heteromorphic regeneration:After regeneration the different organ develops
from lost or removed [Link] phenomenon is also called heteromorphosis.
Example:In Palinurus(shrimp), if the eye is removed from eye stalk then new eye
will [Link] if it is removed along with optic ganglion,antenna like
structure develop in place of eye.
Superregeneration:Development of super numerary organs or parts as a result of
regeneration, termites super [Link]:When incision made in the
planaria head,two head will develop.
Regeneration in Hydra
You already know that the Hydra has spectacular regenerative ability. The hydra is a small
tubular, two layered fresh water animal measuring 20mm in [Link] the Hydra haq a
head or hypostome at the top end, consisting of a mouth,surrounded by a ring of about six large
tentacles which bear a number of stinging cells called [Link] the posterior end, the
hydra has a stalk or penduncle ending in a broad base called pedal or basal disc or foot. The
creature attaches itself to the substratum by means of the pedal disc. Close to the base is the
budding region from where the asexual buds [Link] stomach or gastric region where most of
the digestion occurs is located between the hypostome and the budding [Link] body wall of
Hydra consists of two concentric epithelial layers a) epidermis derived from ectoderm (external
layer) b) gastrodermis derived from the endoderm(internal layer),surrounding a central gastric
[Link] epidermis and gastrodermis are separated by an acellular matrix called mesogle.
Process of regeneration
It has been observed that when the hydra is cut transversely through the gastric [Link]
proximal part gives rise from its distal cut surface to a new hypostome with mouth and tentacles,
and the distal portion regenerates the basal end from its proximal cut surface. Furthermore, it is
also observed that if the hydra is transversely sliced into annular segments, then each segment
gives rise to a hypostome distally (anteriorly )and a basal part proximally (posteriorly).
Sometimes however, it has been noticed that'a short segment, cut just below the hypostome, will
regenerate a hypostome with mouth and ring of tentacles at both ends,
giving rise to a bipolar [Link] in the normal course of life%ydroid polyps are in a
'continuous state of regeneration'.The cells comprising the tentacles and hypostome and the
basal pedal disc get continuously worn out. discarded and replaced by cells shifting distally and
proximally from an area called the growth zone, located below the ring of [Link] growth
zone is easily identified by the presence of large number of interstitial [Link] Hydra, thus there
is a steady migration of cells from the middle of the body in both direction to the hypostome and
tentacles and to the penduncle and [Link] interstitial cells are small undifferentiated cells with
basophilic cytoplasm and relatively large nucleus. These interstitial cells act as a pool of
undifferentiated cells and are capable of being transformed into all the various cell types
(epidermal, endodermal,,nerve cells, gem cells, cnidoblasts etc). Maintenance of life of hydra is
effected by continuos supply of undifferentiatedi nterstitial cells from the growth zone to replace
the worn out and discarded cells at the basal as well as the hypostomal and tentacular [Link]
process of regeneration is 'morphallactic' and blastema formation does not [Link]
process begins with the closure of the wound by contraction of the epitheliomuscular cells of the
epidermis. After this the damaged surface is covered by ectoderm which forms a sheet of
epithelial cells. In the next few hours more than the usual number of cells are given off from the
growth zone towards the cut surface of the damaged area which may be towards the oral or the
aboral side of the [Link] of each piece will occur to form complete miniature hydras
even in complete absence of cell division in either piece.
Source of cells for regeneration
It had been thought earlier that interstitial cells constitute a reserve of undifferentiated cells
which provide the cellular material for regeneration in hydra by mitotic divisions. However
recent studies have shown that cell division in the interstitial cells can be prevented by nitrogen
mustard or irradiations, and still regeneration [Link] cells then taking part are the
differentiated cells of epidermis and gasuodermis which, following amputational injury lose their
specialized features, dedifferentiate and are then reorganized or moulded to from the missing
structures, the hypostome mouth, tentacles or the pedal disc as the case may [Link] cells
are not essential for regeneration, which occurs even when they are put out of [Link]
normally,the interstitial cells also participate in morphallactic regeneration in Hydra. The
ectodermal and endodermal cells belong to two other self-renewing lineages,in hydras like
Pelmarohydra oligaclis and Hydra viridis,the endoderm alone can regenerate an entire animal.
In this type of regeneration, the piece of endodermal cells of hydra first lose their differentiated
features and then redifferentiate into ectoderm, insterstitial cells and endodennal components of
the regenerated part.
Problem of Polarity:The conml of polarity in regenerating coelentrates such as hydra has
received much attention for many years. You are already aware that when a hydra is cut in half,
the half with the basal disc will form a hew hypostome and the half containing the hypostome
will generate a new basal disc. Furthermore, if a hydra is cut into several segments perpendicular
to the body axis, then each middle segment will regenerate both a basal disc and a hypostome.
Thus every region of the hydra can give rise to a new organism .Yet hypostomes, as well foot do
not form anywhere or at all levels along the longitudinal axis of the animal; they form only at the
distal [Link] indicates that a series of gradients arise from the two poles in Hydra which is
rigidly polarized along the distal proximal axis. Grafting experiments have provided further
evidence for the existence of gradients in the hydra. When the hypostome tissue was added to the
middle region of another hydra it formed a new bud with hypostome extending outward. When
the basal disc cells were similarly grafted the new bud extended to form a new pedal disc and
foot. Funhermore if grafts of both basal disc and hypostome cells were grafted together to the
gastric region no supernumerary parts formed. This observation suggests that signals from
opposite ends of the hydra tend to counteract each other, causing the hydra to lose its polarity.
Other experiments have also shown that normal regeneration of the hypostome can be Inhibited
when an intact hypostome is grafted into the body of the hydraof the body,where they
differentiate into appropriate cell types. These experiments and other studies have resulted in the
identification of two opposing sets of gradients arising from the two opposing poles of Hydra-(1)
Head activating and head inhibiting gradients (2) Foot activating and foot inhibiting gradients.
Morphogenetic signalling in hydra is apparently the function of substances of relatively small
molecular weights that are produced at the opposite ends of the [Link] far four distinct
morphogenetic substances of relatively small molecular weight have been isolated from hydra,
which affect regenerative gradient when added to hydra culture medium in micromolar
quantities. The first and the best known head activator is a peptide which accelerate regeneration
of hypostome and tentacles and promotes budding. It has been observed that annular segments of
hydra placed in culture medium containing this activator give rise to bipolar regenerates, and
other strange forms. This is because the polarity is disturbed by the activator. The distribution of
the activator in the hydra body parallels the distribution of nerve cells which are more abundant
in hypostome and less proximally, except for a slight increase in basal region.
LIMB REGENERATION IN AMPHIBIANS
In vertebrates, the amphibians, in particular the urodeles have spectacular power of
regeneration. This power of regeneration has made them a favourite subject of
research and so these animals have been exhaustively studied. Much information
about the processes, mechanisms and systemic factors involved in regeneration in
these vertebrates is now available. We have picked two well studied [Link]
regeneration in vertebrates namely: (i) regeneration of limb in amphibians particularly
in the anurans and (ii) regeneration of lens in the urodele
(i)Sequence of events in urodele limb regeneration:Unlike reptiles, birds and mammals the
urodele amphibians possess unusual ability to regenerate amputated limbs throughout life. The
anuran amphibians (frogs and toads)can also replace the lost part of a limb but only during the
larval or tadpole [Link] all phenomena associated with development, especially those
associated with vertebrate embryos and particularly with the ontogenetic development of limbs
in vertebrates, are inherent in regeneration of limb in [Link] addition,the principles that
regulate patterning of the regenerating limb may be the same as those involved in pattern
formation during the initial development of the [Link] regeneration in the larval and adult
newts has been extensively studied by many investigators, and is by allaccounts the most
exhaustively studied example of epimorphosis. Let us see how the regeneration of limb occurs.
Afier the limb is cut in urodeles, the healing of the wound begins. The epidermis spreads from
edges of the wound to cover the open wound [Link] closure of the wound is a fairly rapid
process and is accomplished in a day or two, depending on the size of the animal. Once this
wound is closed the epidermal cells proliferate an a multilayered mass of cells which form a
conical butge at the tip of the limb stu structure is called the apical epidermal cap (AEC).
Wound healing is accompanied by removal of the debris of damaged and dying cells of the
stump tissues injured by amputation. This debris is removed by phagocytes that accumulate
under the wound epithelium and cause some inflammation for some [Link], many
undifferentiated and morphologically similar mesenchymal cells with large nuclei and basoplilic
cytoplasm accumulate beneath the AEC and give rise to the regeneration bud or [Link]
blastema grows by rapid mitotic divisions of its cells and assumes a more or less conical shape
with the apical epidennal cap at its apex and then enters the phase of redifferentiation.
Redifferentiation to form the missing parts begins with the blastema first becoming spatula
shaped. This is followed by gradual morphogenesis so that the blastema assumes the shape of the
lost parts of the [Link] small regenerated portion then undergoes rapid growth and in course of
time becomes indistinguishable in form and function from the original part that was removed by
amputation as the internal tissues (skeletal elements, muscles, connective tissues, blood vessels
etc.) differentiate from blastema cells. Rudiments of the cartilaginous skeletal elements are the
first tissue to appear as the blastema enters the phase of [Link] various skeletal
elements appear proximo distally just as they do in normal embryonic development of the limb.
The cartiligenous skeletal elements undergo ossification later. Muscles are reformed both by de
novo from blastema cells and by repair of the persisting muscles in the region of [Link]
vessels are not apparent in the very early stages of regeneration but they soon extend into the
blastema from the stump, and in the final regenerated limb the original pattern of vascularisation
gets replicated. Several nerves are also cut when the limb is severed by amputation. However,
soon after amputation their axons grow into the wound and reform the original nerve pattern. As
you will learn later, the nerves play a very important role in limb regeneration. The process of
differentiation, tissue organization and morphogenesis during regeneration are similar to those
during embryonic development. Limb regeneration occurs according to the same sequence of
stages and events (as described above) irrespective of the level of amputation or whether it is the
forelimb or hind limb in the larvae or adult urodeles as also in the tadpoles of anuran amphibians.
Origin of regeneration cells of blastema .The origin of the cells forming the blastema in
vertebrates has been investigated by several workers in the case of regenerating legs of newts
and salamandars. Their studies have shown that cells which form the blastema are of local rather
than of systemic origin. To come to this conclusion they have performed several irradiation
experiments on urodeles. It has been observed that appropriate dose of x-rays supress;he ability
of urodeles to regenerate. Using this knowledge scientists found that those urodeles whose
bodies were irradiated failed to regenerate their limbs after amputation. Likewise, irradiated
limbs also failed to regenerate upon subsequent amputation. However if the limb was shielded
from the x-rays while the rest of the body was irradiated then regeneration proceeded normally in
the limb when the limb was amputated. If only a portion of limb, for example the knee joint, was
irradiated,then regeneration of the limb took place only if the amputation was done above or
below the irradiated segment and not through [Link] problem of potency of blastema [Link]
now know that local cells arc responsible for [Link] was observed that the limb which
regenerated also had its skeletal component. This clearly indicated that skeletal elements can
differentiate from a blastema that received no contribution from preexisting [Link]
tissues, in particular skeletal ones, do not need to be formed by cells from their spccific tissue
counterparts in the [Link] experiment also shows that regeneration blastema arise by the
dedifferentiation of stump tissues. Studies with the help of radioactive tracers have also
positively proved that dedifferentiating tissues are actually involved in the formation of the
blastema and not in one particular cell type. Tissues of amputated limb of newts were supplied
with tritiated thymidine (a DNA precursor) in one experiment and tritiated leucine in another. In
both experiments it was seen that the radioactively tagged precursors were taken up by cells
adjoining the cut surrace. The intake started while the tissue had no!yet lost their histological
characteristics and it was possible to ascertain that all kinds of tissues particularly tlie muscles,
fibroblasts, periosteum, endosteum, Schwann cells of the nerves and thc epictcrmis participated
in the upsurge of synthetic activity. 'The blastema was later found to consist of labelled cells of
all these tissues. It follows Regeneration thus that none of the tissues of the amputation stump are
excluded from participation in regeneration. But that all cells respond to the wounding by growth
(synthesis) and later by proliferation. However, it has been experimentally observed that
epidermis does not contribute to the internal [Link], biologists alternatively also
wonder if there exist local population of reserve cells that retain the embryonic property of
pluripotency and so are capable of forming the various differentiated tissues in the limb
regeneration. They have found it extremely difficult to test this alternative hypothesis
experimentally, as the limb tissues are not made of pure populations of a single cells type.
Furthermore, in addition to specialised cells most tissues have connective tissue cells some of
which may be pluripotent.

The standard procedure used by investigators for examining or finding out the differentiation
potential of tissues is to irradiate a diploid host animal. Irradiation prevents proliferation of the
host cells, thus limiting their ability to take part in regeneration. After irradiation a tissue implant
is made in the host from a uiploid donor. The Iimb is then severed in order to allow it to
regenerate. After regeneration the regenerate is subjected to histological analysis. The triploid
nuclei in the tissues indicate that they are from the donor tissue, and if found in the regenerate-as
they usually are, they indicate that they have participated in the regeneration of new tissues.
Often in some experiments the donor triploid cells are labelled with [Link] donor
cells due to the presence of the triploid radioactive nuclei, are easily [Link] has
experimentally been observed that when donor tissue is cartilage from which the muscle and
connective tissues have been carefully cleaned, then the regenerate has cells of donor type in the
regenerated cartilage, namely perichondrium (the connective tissue layer surrounding the
cartilage) connective tissue of joints and fibroblasts' but,no donor cells are found in the epidermis
or muscle. However, it is reported that the cartilage taken from limb,of young axolotl larvae but
not the adult implanted into irradiated limb of host axolotl contributed cells to some muscle
tissue .
regenerate which developed after amputation of host limb through the implant. On the other hand
several investigators have found that if muscle is the donor tissue then the donor cells of muscle
origin are found in all regenerated mesodermal derivatives including that cartilage. This result
naturally suggests that the dedifferentiated cells originating in muscle tissue have greater potency
than the cells derived from cartilage tissue. However, you should know that muscle is intimately
associated with perimuscular fibroblasts which cannot be rigorously excluded, and could be the
source for differentiation of tissues other than muscle in the regenerate in which muscle from a
donor was [Link] in one series of such experiments on larval Xenopus, cell bearing
a nucleolar marker cloned from single myoblasts or fibroblasts were implanted into limb of a
different host Xenopus, which were later amputated. It was found that cells of both myoblast and
fibroblast origin can form all the various mesodermal tissues in the [Link] is
incapable of producing mesodermal tissues and it definitely does not contribute any cells to the
blastema, However, skin dermis contributes cells of fibroblast nature which can differentiate into
several mesodermal [Link] studies on the axolotl. (Ambystoma americanum)
using diploid and triploid cell markings have revealed that a very large proportion (43%) of the
blastemal cells are of dermal fibroblasts origin. Both dermis and muscle tissue contain a larger
number of fibroblasts. Therefore, there does exist the likelihood that many of the mesodermal
derivatives of the regenerate are produced by fibroblast [Link] of wound epidermis and
'Apical Epidermal Cap'You already know; that following amputation, the stump epidermal cells
at the wound edge migrate over and rapidly cover the wound to from a multilayered apical
epidermal cap. The formation of this cap depends on the carly innervation of the wound
epidermis by nerve fibres. The cap fails to form if the limb is denervated before or immediately
after amputation. If this cap is removed regeneration fails to occur. If an additional cap is grafted
on the developing blastema it induces regeneration of supernumerary limb. Thus the apical
epidermal cap formed by wound epidermis is necessary for permitting regeneration. Any other
epidermis grafted on the wound surface does not form the apical cap and does not support
[Link] shows that only the apical cap which developed from the wound epithelium
could promote and support regeneration of the limb. In the non-regenerating amputated limb-
stumps of older tadpoles and adults of frogs, the characteristic apical epidermal cap does not
develop and so regeneration does not [Link] results indicate that wound epidermis
stimulates dedifferentiation and mitosis in the underlying cells arising from stump tissue. thus
inducing the formation and growth of the blastema and the subsequent regeneration of the limb.
The blastema cells in zone immediately adjacent to the apical cap remain in an undifferentiated
and proliferating state, while those proximal to this zone begin to differentiate. In this way
proximal distal pattern of redifferentiation of blastema cells is controlled by the epidermal cap.
You may recall that the apical ectodermal ridge (AER)of the embryonic limb bud also plays a
similar role in allowing distal outgrowth of the limb. Both the AER in,embryonic limb
development and the apical cap of the regenerating limb stimulate cellular proliferation in cells
located beneath them and keep them from differentiating,which enables differentiation of limb
tissues in a proximo-distal sequence.
Role of Nerves:It has been observed that soon after amputation nerves invade the regeneration
blastema. If the stump is denervated by cutting the nerves supplying the limb as they emerge
from the spinal cord, and are prevented from regrowing into the site of amputation them
regeneration fails to occur. The apical cap is not formed. In larval urodeles, denervation results in
large scale regression of stump tissues and the entire limb stump may [Link] the nerve
fibers are allowed to regrow into the stump before a thick skin forms on the wound surface
regeneration may be reintiated. These studies have shown with certainty that the presence of
nerves is essential for regeneration of the limb. A number of experimental studies have been
made to understand the nature and mechanism of nerve influence on limb regeneration in
urodeles. The nerves are believed to produce a "trophic influence" on the blastema cells of the
regenerating [Link] neurotrophic effect has been found to be produced by all nerves, whether
motor or sensory or central. irrespective of whether they have functional association with the
central nervous system or not. Grafting of spinal ganglia in the regenerating area of limbs, whose
own nerves have been cut and severed, also promote [Link], it has been found
that the presence of a minimum number of nerve fibers is necessary for regeneration. Below this
threshold regeneration does not take [Link] have shown that the neurotrophic
influence is essential for initiating regeneration and for the growth of the blastema. But once the
blastema begins redifferentiation and morphogenesis the subsequent course of regeneration is
independent of the nerves. Denervation at this stage does not prevent completion of
regeneration [Link] has been observed that vertebrate species (such as advanced tadpoles and
adults of anurans), in which limb regeneration does not occur, contain fewer nerve fibres per unit
area of amputation wound as compared to the urodele Triturus. It is possible that inability of
these species to regenerate a limb may at least be partially due to an inadequate nerve supply.
This hypothesis finds support from the results of experiments in which growth of regeneration
blastema was induced in the amputated limbs of lizards, frogs and the marsupial opposum by
increasing the nerve supply at the site of [Link] have demonstrated that nerves
exert their effect on regeneration of limbs by promoting mitosis of blastema cells and synthesis
of DNA and proteins Recent studies have shown that the neurotrophic factor is probably a
fibroblast growth factor(FGF), a protein with a molecular weight of about 13,000. It is released
from myelin,the chief component of nerve sheaths. FGF is present in the nerve extracts and is
also present in the blastema. It has been found to stimulate m'itotic activity. in the blastema of
denervated limbs. Role of Hormones in [Link] neurosecretory effects in
regeneration are integrated into a neuroendocrine feed back system. It is often difficult to
distinguish hormonal from neurai influences during regeneration, particularly in invertebrates,
although the two influences are easily distinguished in [Link] amphibians, the hormones
of the three glands, namely pituitary, adrenal and thyroid appear to affect limb regeneration.
These hormones control different phases of the regeneration process though the mechanism of
their action is till not very [Link] the hormonal response varies with the age of
animal. For example,larval urodeles regenerate limb in total absence of pituitary hormone,
whereas the adult is completely dependent on the pituitary gland for limb regeneration, a
dependence acquired during metamorphosis. The role of the pituitary gland and its hormones in
regulating regeneration has been worked out with great difficulty by mean of several experiment.
In some experiments the pituitary gland of the newt was removed (hypophysectomy)at the time
of limb amputation while in others it was removed later. Results showed that hypophysectomy at
the time of amputation inhibited regeneration whilc delayed hypophysectomy until several days
after amputation allowed some regeneration; to occur. The extent of regeneration was found to
be dependent on the delay between amputation and operation. Hyposectomy performed three
days after amputation allowed some limb regeneration, while that performed after 13 days or
more allowed the entire limb to regenerate. These findings indicated that pituitary hormones are
needed only during the very early stages of regeneration (like wound healing).Other studies
indicate that pituitary glands exert only an indirect effect on regeneration by stimulating the
adrenal gland to produce cortisone. This observation is based on two experiments. In one
experiment the regenerative capacities of the hypophysectomized newts were restored by
replacement therapy of either cortisone(secreted by adrenal gland) or adrenocorticotropic
hormones (ACTH) (secreted by pituitary). In the other experiment it was found that inhibition of
cortisone secretion by administration of drugs, inhibited regeneration, which could again be
relieved by cortisone but not by ACTH of the pituitary. These experiments thus indicate that the
pituitary gland is regulated by the adrenals. The role of cortisone has also been found to be
apparently limited to early phase of wound healing as the formation of blastema appears to be
independent of the presence of cortisone which only promotes wound healing. In the absence of
cortisone. In the absence, of cortisone wound healing fails and a thick dermal pad forms at the
stump, preventing [Link] is produced by thyroid gland. It affects regeneration
and also controls metamorphosis in amphibians particularly in anurans where the larval adult
transformation is most dramatic .The effect of thyroxine in regeneration is however poorly
understood as it inhibits tadpole limb regeneration if administered before amputation, but
accelerates morphogenesis if given at the blastemal stages. Thyroxine inhibition is consisterft
with loss of regenerative capacity after a larva transforms into an adult. Thyroxine stimulates
limb morphogenesis in the anuran tadpole but once formed, a limb loses thyroxin dependency
and its regeneration is inhibited by thyroxin [Link] of distal Transformation of Blastema:
An intriguing phenomenon characteristic of limb regeneration is, that only the part of the limb
removed distal to the level of amputation is regenerated. For example,suppose a fore limb is cut
through the middle of upper arm, removing the distal half of the upper arm, wrist and hand
leaving behind a stump consisting of only the proximal half of the upper arm. In this case the
blastema formed at the cut end of the stump produces a regenerate consisting of the distal part of
upper arm, lower arm,wrist and hand in this order; the proximal part of the upper arm is not
duplicated in the [Link] a complete fore limb is regenerated without any duplication.
The blastema always forms siructures or parts distal to its own level of origin, along the
proximo-distal axis of ~ h clim b. Fur~hcrmorer, egardless of the level of amputation the
blastema formed always regenerates only the distal structures, even if the polarity of stump is
reversed as demonstrated in an elegant experiment on axolotl [Link] is known as the Rule of
Distal Transformation of Blastema and applies equally to limb regeneration in urodeles and
anurans amphibians as well as to leg regeneration in insects According to the recently
propounded theory of positional information based on the concept of gradients, it is believed that
cells at various levels along the proximo-distal(P-D)a xis of the limb have the information
specifying their position at that [Link] have been called positional values. For example,
suppose the cells at various levels of a fprelimb along P-D axis are considered to have positional
values, say from 1 to 10 with 1 specifying the position at the base near the girdle and 10 the
position at the top of the digits and position 5 falling below the elbow in the lower arm. If
amputation is made across the level 5, the cells of blastema formed at this level are able to
generate the missing positional values 6 to 10 i.e. distal to the level of amputation but not the
values 1-5 proximal to that level. Only the missing positional values are supposed to be
regenerated in the blastema. The formation of parts only distal to the level of amputation is thus
sought to be explained in terms of positional values along the proximo distal axis or the limb.
Experiments have also indicated that the mechanisms which regulate regeneration in tetrapod
limbs may also, be the same which regulate pattern formation during development. It has been
experimentally secn that not only does regeneration mimic embryonic limb development, but
both regenerating and developing limb tissues can also interact with each other to form a normal
[Link], supernumerary digits resulted if the axis of the regenerating blastema and stump
were [Link] is hoped that studies on developing and regeneration systems may explain the
control of pattern in both cases, and furthermore outline, common principles in pattern formation
in other developing systems. It has been found, however, that the rule of distal transformation of
blastema can be subverted if the amputated limbs of urodeles and anuran tadpoles are treated
with a suitable amount of any of the derivatives of vitamin A, collectively known as retinoids,
including vitamin A palmitate, Vitamin A alcohol, retinoic acid [Link] 1970s while studying
regeneration of amputated limbs in anuran tadpoles [Link] and his coworkers at the Zoology
Department of Rajasthan University, Jaipur,found that when vitamin A palmitate (a retinoid) was
added to the water in which the tadpoles were kept, the regenerated part formed was not only the
distal part that had been removed by amputation; instead, in many cases complete limbs
regenerated consisting of pans both distal as well as proximal to the level of amputation. This
showed that the restriction on the blastema to form only distal structures was removed [Link]
retinoid. In many cases more than one such limb (mirror images of each other)regenerated from
the same stump. This was the case at whatever level the tadpole limb was amputated It has been
found, however, that the rule of distal transformation of blastema can be subverted if the
amputated limbs of urodeles and anuran tadpoles are treated with a suitable amount of any of the
derivatives of vitamin A, collectively known as retinoids, including vitamin A palmitate, Vitamin
A alcohol, retinoic acid [Link] 1970s while studying regeneration of amputated limbs in
anuran tadpoles [Link] and his coworkers at the Zoology Department of Rajasthan University,
Jaipur,found that when vitamin A palmitate (a retinoid) was added to the water in which the
tadpoles were kept, the regenerated part formed was not only the distal part that had been
removed by amputation; instead, in many cases complete limbs regenerated consisting of pans
both distal as well as proximal to the level of amputation. This showed that the restriction on the
blastema to form only distal structures was removed [Link] retinoid. In many cases more than
one such limb (mirror images of each other)regenerated from the same stump. This was the case
at whatever level the tadpole limb was amputated This dramatic effect of retinoids on the pattern
of limb regeneration indicates that the retinoids apparently resets the positional information of
the cells in the blastema to amore ~roximalv alue so that the cells that would form distal
structures arereproirammed to f ? n proximal structures as well. In other words, the genetic-D ro-
n rarnme or the develo~mentawl tential of the blastema cells involved'in regeneration is changed
by the rehoids. Thiseffect is now referred to as proximalization of the blastema
(ii)Limb regeneration in anuran amphibians:You are already aware that the limb of adult
frogs do not regenerate after amputation. However some regeneration has been found to occur in
experimental animals. Recovery of regeneration ability in most metamorphic frogs is made
possible by tissue trauma and prevention of wound [Link] is achieved by extensive
piercing of the amphibian amputaton site with a needle as well as traumatization by using
hypertonic solutions. Normally wound healing in adult anurans is brought about by the
movement of the epidermis and the dermis over the wound surface. In contrast only the
epidermis covers the wound in the urodeles. Normally whcn a part of the adult anuran is
amputated the wound heals by the production of connective tissue from dermal elements
(derma1ization) and the scarring of injured [Link] injury in some way interferes with
dermalization and allows a variable amount of regeneration to occur. Similar results of
regeneration have been obtained by implanting batteries and applying a continuous'direct current
through fine wire to the amputation area. Superficially this appears to stimulate action of the
nerves. Regeneration also sometimes results from irritation. Implanting of adrenal glands also
appears to prevent [Link] has been observed that if larval skin is applied over adult
frogs then regeneration becomes possible. Other studies have implicated the wound epithelium
as an important factor in initiating regencration.
 PARTHENOGENESIS (VIRGIN ORIGIN)
Usually an unfertilized ovum develops in to a new individual only after the union with the
sperm or fertilization but in certain cases the development of the egg takes place without the
fertilization. The peculiar mode of sexual reproduction in which egg development occurs
without the fertilization is known as the [Link] an un-fertilised ovum
develops into a new individual only after the union with the sperm or fertilisation but in certain
cases the development of the egg takes place without the [Link] peculiar mode of
sexual reproduction in which egg development occurs without fertilization is known as parthen
-ogenesis (Gr., parthenos = virgin; genesis = origin).The phenomenon of parthenogenesis occurs
in different groups of the animals as in certain insects (Hymenoptera, Homoptera,Coleoptera),
Types of Parthenogenesis:
The parthenogenesis may be of two types: 1. Natural parthenogenesis 2. Artificial parthenogenesis
1. Natural Parthenogenesis:In certain animals the parthenogenesis occurs regularly,
constantly and naturally in their life cycles and is known as the natural [Link]
natural parthenogenesis may be of two types, viz., complete or incomplete:
(i) Complete Parthenogenesis: Certain insects have no sexual phase and no males. They depend
exclusively on the parthenogenesis for the [Link] type of parthenogenesis is known
as the complete parthenogenesis or obligatory parthenogenesis. E.g. Lacerta saxicola
(ii) Incomplete Parthenogenesis:- The life cycle of certain insects includes two generations the
sexual generation and parthenogenetic generation, both of which alternate to each other. In such
cases, the diploid eggs produce females and the unfertilized eggs produce males. This type of
parthenogenesis is known as the partial or incomplete or cyclic [Link] life cycle
of certain insects includes two generations, the sexual generation & parthenogenetic generation,
both of which alternate to each other. In such cases, the diploid eggs produce females and the un-
fertilised eggs produce males. This type of parthenogenesis is known as the partial or incomplete
or cyclic parthenogenesis. The complete or incomplete type of natural parthenogenesis may be
of two types: 1. Haploid /arrhenokous parthenogenesis 2. Diploid /thelytokous parthenogenesis
1. Haploid or arrhenotokous parthenogenesis: In the arrhenotokous parthenogenesis, the
haploid eggs are not fertilised by the sperms and develop into the haploid individuals. In these cases,
the haploid individuals are always males and the diploid individuals are the females. e.g.
a. Insects: (i) Hymenoptera (bees and wasps), (ii) Homoptera, (iii) Coleoptera (Micromalthus
debilis), (iv) Thysanoptera (Anthothrips verbasi).
b. Arachnids: Arachnids, e.g., ticks, mites and certain spiders (Pediculoides ventricusm),
c. Rotifers: Rotifers, e.g., Asplanchne amphora
2. Diploid or thelytokous parthenogenesis:- In the diploid parthenogenesis, the young
individuals develop from the unfertilized diploid eggs. Following types of the thelytoky have
been recognized. (i) A meiotic parthenogenesis:- Sometimes during the oogenesis, first meiotic
or reduction division does not occur but second meiotic division occurs as usual. Such eggs
contain diploid number of chromosomes and develop into new individuals without the
fertilization. This type of parthenogenesis is known as apomictic or ameiotic parthenogenesis
and occurs in Trichoniscus (Isopoda), Daphnia pulex (Crustacea), Campelona rufum (Mollusca),
Weevils and long-horned grasshoppers(ii) Meiotic parthenogenesis:- Certain eggs develops by
the usual process of oogenesis but at certain stages diplosis or doubling of chromosome number
and production of diploid eggs occur. Such eggs develop into the diploid individuals and this
phenomenon is known as the meiotic parthenogenesis. The diplosis of the diploid thelytoky
may occur by the following methods:- (i) By autofertilization:- In certain cases the oocyte
divides meiotically up to the formation of ootid or ovum and secondary polocyte. But the ootid
and the secondary polocyte unite together to form a diploid egg which develops into a new
individual, e.g., Artemia salina (Crustacea) and various other organisms. (ii) By restitution:-
Sometimes in primary oocyte karyokinesis forms a nucleus of the secondary oocyte and nucleus
of the first polocyte. The chromosomes of both daughter nuclear arranged on the equator and
undergo second meiotic division to form a diploid ootid and a diploid polocyte. The diploid
ootid or ovum develop into a parthenogenetic diploid individual. This type of diplosis is known
as the restitution, e.g., insects of order Hymenoptera (Nemertis conesceus) and LepidopteraIn
the diploid parthenogenesis, the young individuals develop from the unfertilised diploid eggs.
Following types of the thelytoky have been recognised:
(i) Ameiotic Parthenogenesis: apomixis (apomicitic parthenogenesis)
Sometimes during the oogenesis, first meiotic or reduction division does not occur but second
meiotic division occurs as usual. Such eggs contain diploid number of chromosomes and develop
into new individuals without the fertilisation. This type of parthenogenesis is known as apomictic
or ameiotic parthenogenesis and occurs in Trichoniscus (Isopoda), Daphnia pulex (Crustacea),
Campelona rufum (Mollusca), weevils and long-horned grasshoppers.
(ii) Meiotic Parthenogenesis: Automixis (automictic parthenogenesis)
Certain eggs develop by the usual process of oogenesis but at certain stages diplosis or doubling
of chromosome number and production of diploid eggs occur. Such eggs develop into the diploid
individuals and this phenomenon is known as the meiotic parthenogenesis.
The diplosis of the diploid thelytoky may occur by the following methods:
(i)By Autofertiiisation: In certain cases, oocyte divides meiotically up to formation of ootid
/ovum & secondary polocyte. But the ootid & secondary polocyte unite together to form a
diploid egg which develops into a new individual, e.g., Artemia salina (Crustacea) and various
other organisms.

(ii) By Restitution:
Sometimes in primary oocyte, karyokinesis forms a nucleus of the secondary oocyte and nucleus
of the first polocyte. But the karyokinesis is not followed by the cytokinesis. The chromosomes
of both daughter nuclei are arranged on the equator and undergo second meiotic division to form
a diploid ootid and a diploid polocyte. The diploid ootid or ovum develops into a parthenogenetic
diploid individual. This type of diplosis is known as the restitution, e.g., insects of order
Hymenoptera (Nemertis conesceus) and Lepidoptera.
Parthenogenesis in order Hymenoptera
In the insect order Hymenoptera (which includes bees, wasps, and ants), parthenogenesis can
take one of three forms: arrhenotoky, thelytoky, and deuterotoky. In arrhenotoky, haploid males
are produced from unfertilized eggs laid by mated (impregnated) females or by so-called
secondary, or supplementary, queens, which have not been impregnated. In thelytoky, which
occurs in many species of the suborder Symphyta (a group that includes the sawfliesthe
horntails, and the wood wasps), unmated females produce males. In deuterotoky, unmated
females of some Symphyta produce females as well as males. The occurrence of these forms is
not always mutually exclusive. For example, in Apis (bees), about 1 percent of the eggs laid by
secondary queens may be [Link] associated with arrhenotoky, thelytoky, and
deuterotoky is pseudoarrhenotoky (or paternal genome elimination). Pseudoarrhenotoky is a
non parthenogenic form of reproduction that occurs in the hymenopteran superfamily
Chalcidoidea (a group of small parasitic wasps) and in some mites, Like arrhenotoky,
pseudoarrhenotoky results in the production of haploid males. In this process, development
begins as diploid organisms within fertilized eggs; however, as development progresses, males
become haploid after the paternal contribution to genome has been lost,eliminated, or deactivated
2)Artificial Parthenogenesis: The eggs which always develop into the young individuals
by the fertilisation sometimes may develop parthenogenetically under certain artificial
conditions. This type of parthenogenesis is known as artificial parthenogenesis.
.
The artificial parthenogenesis may be induced by various chemical and physical means.
A. Physical means:The following physical means cause the parthenogenesis:
(i) Temperature the range of temperature may induce parthenogenesis in the eggs. For instance,
when the egg is transferred from the 30°C to 0-10°C, the parthenogenesis is induced(ii)Electrical
shocks can cause parthenogenesis(iii) Ultraviolet light can cause parthenogenesis(iv) When eggs
are pricked by fine glass needles the development of young ones takes place parthenogenetically.
B. Chemical means:following chemicals have been found to cause parthenogenesis in
normal eggs: 1. Chloroform; 2. Strychuine; 3. Hypertonic and Hypotonic sea waters;
4. Chlorides of K+, Ca++, Na+, Mg++, etc.; 5. Acids such as butyric acid, lactic acid, oleic acid
and other fatty acids; 6. Fat solvents, e.g., toluene, alcohol, benzene and acetone7. Urea and
sucrose. The artificial parthenogenesis has been induced by above mentioned physical and
chemical means by various workers in the eggs of most echinoderms, molluscs, annelids,
amphibians, birds and mammals.
Significance of Parthenogenesis
1. The parthenogenesis serves as the means for the determination of sex in the honeybees, wasps,
2. The parthenogenesis supports the chromosome theory of inheritance 3. The parthenogenesis is
the most simple, stable and easy process of reproduction 4. The parthenogenesis eliminates the
variation from the populations. 5. The parthenogenesis is the best way of high rate of
multiplication in certain insects, e.g., aphids. 6. The parthenogenesis causes the polyploidy in the
organisms 7. The parthenogenesis encourages development of the advantageous mutant
characters 8. The parthenogenesis checks the non-adaptive combination of genes which may be
caused due to the mutation 9. Due to the parthenogenesis, there is no need for the organisms to
waste their energy in the process of mating but it allows them to utilise that amount of energy in
the feeding and reproduction. 10. The parthenogenesis avoids the sterility in the races.
However, parthenogenetic forms, i.e., individuals produced due to parthenogenesis are not much
successful in the struggle for existence due to the fact that no recombination of genetic material
occurs, hence, variations are not produced.
 Honey bee as an example of parthenogenesis:In honey bees, parthenogenesis is the
asexual reproduction of an unfertilized egg, leading to the development of haploid male
offspring (drones). A female honey bee (the queen or a worker) produces an unfertilized
egg, which then develops directly into a male bee without fertilization by a sperm. This
process is called arrhenotokous parthenogenesis and is a form of haplodiploidy, where
males have one set of chromosomes (haploid) and females have two (diploid).

1. Egg Production: The queen bee produces eggs, some of which are fertilized
and others are not. 2. Unfertilized Egg Development: An unfertilized egg
begins to develop through parthenogenesis. 3. Male Offspring (Drones):The
unfertilized egg develops into a haploid drone (male), receiving genetic material
only from the queen. 4. Haplodiploidy:This creates a sex determination system
where females are diploid (from fertilized eggs) and males are haploid (from
unfertilized eggs).
2. Key Points

 Asexual Reproduction: Parthenogenesis is a form of asexual reproduction.

 Male Development: It is the normal way male honey bees (drones) are produced.
 Haploid Nature: Drones are haploid, meaning they have half the number of
chromosomes as the females.
 Queenless Colonies: Even queenless worker bees can lay unfertilized eggs that
develop into males.
 Thelytoky: In a specific case (primarily in the South African Cape honey bee), a form of
parthenogenesis called thelytoky can occur, where unfertilized eggs develop into
females
Ageing
Entropy always wins. Each multicellular organism, using energy from the sun, is able to develop
and maintain its identity for only so long. Then deterioration prevails over synthesis, and the
organism ages. Aging can be defined as the time-related deterioration of the physiological
functions necessary for survival and fertility. The characteristics of aging as distinguished from
diseases of aging (such as cancer and heart disease) affect all the individuals of a [Link]
procedure begins after the organism has attained sexual maturity and it ends lastly in the death
of the organism. Most animals undergo the process of ageing, which is also known as
senescence. The branch of study dealing with old age and ageing is called Gerontology. Ageing
varies from a few days to a few years or more. Man has an average life span of 100 years that is
set by a group of death genes. Once the juvenile period, which is a period of active growth under
hormonal influence, is completed and the organism enters the adult phase, growth is replaced
by a metabolic equilibrium between anabolism and catabolism. Some animals tend to escape
ageing by evolving a rejuvenation process. Bryozoans are excellent examples, which get rid off
ageing effects by discarding “brown bodies” and thus are able to rejuvenate a new life. No
amount of improvement in sanitation, richness in diet or improvement of medical care facilities
can increase the longevity of mammals. Such improvements may simply increase the percentage
of survivors at lower ages in the population. The process of ageing is the progressive decline in
all vital activities of the organism that terminate in death goes on at different rates in animals of
various species and in different individuals of the same species. At cellular level, the
following processes are involved: 1. Possible reduction in functional efficiency of non-
dividing highly specialized cells such as nerve cells, muscle cells, etc. 2. Progressive stiffening
with age of the structural protein collagen which constitutes more than a third of all body
proteins and serves as general binding substance of skin, muscular and vascular [Link] is
also the substance of cartilage and tendons, it fills up the spaces between muscle fibres and
between the cells of many organs, serving as the stabilizing fibre of connective tissue. In skin, a
thick mating of collagen fibres gives the skin its toughness and plasticity. During ageing process
collagen accumulates changes in the molecular structure that affects its integrity and function.
The ageing of collagen may provide an objective index for determination of ‘biological age’
asdistinguished from ‘calendar age’ of men and other animals. Once the fibres of collagen are
laid down in the body they cannot be renewed. Collagen molecule consists of 3 strands wound
around to form helices. During collagen ageing cross links increase in number between the
different strands and different molecules. 3. Limitation of cell division:Hayflick (1968) showed
that as population of human fibroblasts approaches the end of its lifetime, aberrations often
crop up in the chromosomes. Chromosome aberrations and cell division peculiarities related to
age have also been observed in leucocytes and in liver. In man several organs lose weight after
middle age, which can be attributed to cell loss. The human brain weighs considerably less in old
age than it does in middle age. Kidney also shows reduction in nephrons accompanying cell loss
and the number of taste buds per papilla of tongue drops from 245 in young adults to 88 in
aged. Animal age may also result from deterioration of the genetic program that orchestrates the
development of cells. As time goes on, the following changes occur. 1. The DNA of dividing cells
may become clouded with an accumulation of copying errors. 2. The coding and decoding
systems that govern the replication of DNA operates with high degree of accuracy, but the
accuracy is not absolute. 3. Certain enzymes involved in the transcription of information from
DNA for the protein synthesis may deteriorate with age. All these events lead to loss of
progression of cells as a result of ageing and eventually to the death of cells. 2. Cellular Changes
during Ageing Ageing causes changes in cells at numerous levels which can be 1. Morphological
changes, 2. Physiological changes and 3. Subcellular changes. 1. Morphological changes: The
most obvious morphological changes occurring during ageing in cells may be as follows: a)
Decline in cell volume: The cell undergoing ageing exhibits reduced cell volume. Reduced cell
volume and cell death results in decreased weight of organs, thereby resulting in progressive
decline of body weight. b) Accumulation of exhaustion pigment: The exhaustion pigment
lipofuscin, yellow pigment or brown degenerations are peroxidation byproducts of unsaturated
membrane lipids (fatty acids) and denatured proteins. These are present in nerve and cardiac
muscle cells and also in other cells of the body to a lesser degree. c) Nuclear Pyknosis: With
advancing age, the nucleus becomes shrunken and stains deeply. Such a nucleus is called
pyknotic and the degenerative process is known as nuclear pyknosis. d) Formation of lipid
vacuoles: Ageing results in accumulation of small lipid vacuoles in the cytoplasm. 2.
Physiological Changes: Various physiological and biochemical changes occurring in the ageing
cells are: a) Collagen and ageing: Collagen, an essential basic protein present in the connective
tissue becomes cross-linked and stiff with ageing. b) Calcium and ageing: In the ageing cells,
Ca++ ions are found to accumulate in the peripheral cytoplasm due to changes in the
permeability of cellular membranes. c) DNA and Cellular ageing: Loss of nuclear material and
DNA damage progresses with age. Due to this, RNA synthesis and protein production also
decline leading to cell loss with age. d) RNA and cellular ageing: There are some evidences of
quantitative age changes in the metabolism of individual RNA species. Decline in transcription
may result from alterations in chromosome organization, specifically at the level of
chromosomal proteins that complex with DNA to form chromatin. e) Protein synthesis and
cellular ageing: The synthesis of protein universally declines with age in all eukaryotic
organisms. Ageing also causes decline in rRNA and alterations in tRNA (required for amino acid
binding).Elongation factors necessary for stabilization and continued elongation of the
polypeptide chain decrease with age. Post-translational modifications also occur during ageing.
Other biochemical changes associated with ageing are1. Increase in cholesterol and triglyceride
level, 2. Increase in Blood globulin levels, 3. Decrease in alkaline and acid phosphatases4.
Decrease in cellular respiration and 5. Increase in serum creatinine 3. Subcellular changes:
Various subcellular changes associated with ageing are: a) Plasma membrane and ageing: The
functional capacity of plasma membrane gets disturbed during ageing. Its permeability changes.
Ca++ becomes accumulated in the membrane. Total lipids of membranes decrease with age. The
cholesterol: phospholipid ratio in cellular membranes increase with age. b) Endoplasmic
reticulum and ageing: The amount of granular E. R. decrease in the cytoplasm of old cells. In the
nerves of older animals and human beings, there is decrease of Nissl granules (rRNA). c)
Mitochondria and ageing: Due to ageing, the rate of carbohydrate metabolism is decreased.
Ageing also affects glycolysis. The mitochondria are involved in the ageing process of the
animals and in old tissues, the mitochondria become degenerated. d) Nucleus and ageing:
Nucleus remains unaltered in ageing cells. The change in nucleo-cytoplasmic ratio acts as an
important index for natural senescence and death. Incidence of loss of one X-chromosome in
woman and Ychromosome in man have been reported in ageing cells. Ageing in Animals:
Animals from protozoans to vertebrates exhibit ageing and senescence. Indefinite life span is
found in those animals which exhibit asexual reproduction and continued growth or
replacement of all body parts throughout life. This is related to genetic constitution of
individuals that plays a role in ageing. In mammals, a relationship also exists between the ratio
of brain weight to body weight and life span and the proportion of life spent in the
prereproductive stage and reproductive phase; the more delayed the onset of reproductive life,
the longer the life span and the later the onset of senescence.
Causes of aging
The general senescent phenotype is characteristic of each species. But what causes it? This
question can be asked at many levels. We will be looking primarily at the cellular level of
organization. Even here, there is evidence for many different theories, and there is not yet a
consensus on what causes aging.
[Link]-and-tear theory:The “wear-and-tear” theory assumes that animals and cells, like
machines, simply wear out. Animals, however, unlike machines, have some ability to repair
themselves, so that this theory does not fit the facts of a biological system. A corollary to the
wear-and-tear theory is the presumption that waste products accumulate within cells and
interfere with function. The accumulation of highly insoluble particles, known as “age pigments,”
has been observed in muscle cells in the heart and nerve cells of humans and other animals.
2. Cross-linking theory:With increasing age, tendons, skin, and even blood vessels lose
elasticity. This is due to the formation of cross-links between or within the molecules of collagen
(a fibrous protein) that give elasticity to these tissues. The “cross-linking” theory of aging
assumes that similar cross-links form in other biologically important molecules, such as
enzymes. These cross-links could alter the structure and shape of the enzyme molecules so
that they are unable to carry out their functions in the cell.
3. Autoimmune theory:Another theory of aging assumes that immune reactions, normally
directed against disease-producing organisms as well as foreign proteins or tissue, begin to
attack cells of the individual’s own body. In other words, the system that produces antibodies
loses its ability to distinguish between “self” and foreign proteins. This “autoimmune” theory of
aging is based on clinical rather than on experimental evidence.
4. Oxidative damage theory:Reactions that take place within cells can result in the oxidation of
proteins & other cellular [Link] entails the loss of electrons from these molecules,
causing them to become unstable and highly reactive and leading to their eventual reaction with
and damage of cell components such as [Link] reactive molecules are known as
free radicals—any atom or molecule that has a single unpaired electron in an outer shell.
Oxidative damage (oxidative stress) accumulates with age, and this has given rise to the free
radical theory of aging, which is concerned in particular with molecules known as reactive
oxygen species (ROS). This theory was first proposed in the 1950s by American gerontologist
Denham Harman and was supported in part by evidence that antioxidant proteins, which
neutralize free radicals, are more abundant in aging cells, indicating a response to oxidative
[Link] initial free radical theory of aging was later extended to include ROS derived from
cellular organelles known as mitochondria, which are the primary sites of energy production in
most eukaryotic organisms (eukaryotic cells are cells with clearly defined nuclei). The
mitochondrial theory of aging was based on the idea that there exists within mitochondria a
vicious oxidation cycle, in which the mutation of mitochondrial DNA impairs the function of
proteins in the organelle’s respiration machinery, thereby enhancing the production of DNA-
damaging oxygen radicals. This in turn results in the accumulation of mutations in mitochondrial
DNA and a bioenergetic impairment, characterized by the failure of mitochondria to produce
sufficient energy for cells to carry out their daily activities, which leads to tissue dysfunction and
degeneration. A similar mitochondrial theory of aging proposes a mechanism in which electrons
leaking from the electron transport chain (ETC), the central component of the organelle’s
respiration machinery, produce ROS and then damage ETC proteins and mitochondrial DNA,
leading to further increases in intracellular ROS levels and a decline in mitochondrial function.
Another consideration is the molecular inflammatory theory of aging, whereby the activation of
redox- (oxidation-reduction-) sensitive transcription factors (molecules that control gene activity)
by age-related oxidative stress causes increased expression of proinflammatory genes, leading
to inflammation in various tissues. This inflammatory cascade is exaggerated during aging and
has been linked to many age-associated pathologies, including cancer, cardiovascular disease,
arthritis, and several neurodegenerative diseases. Chronic inflammation, whether due to diet,
infection, stress, or other factors, can potentially accelerate the aging [Link] under
calorie restriction produce fewer ROS and age slower. Such effects of calorie restriction have
been attributed to its ability to lower the steady state of oxidative stress, slow the accumulation
of age-associated oxidative damage, and increase metabolic efficiency.A common phenomenon
in all of the aforementioned theories is that ROS serve as a contributing factor to many age-
associated diseases.
5. Mitochondrial genome damage:The mutation rate in mitochondria is 10–20 times faster
than the nuclear DNA mutation rate (Johnson et al. 1999). It is thought that mutations in
mitochondria could (1) lead to defects in energy production, (2) lead to the production of ROS
by faulty electron transport, and/or (3) induce apoptosis. Age-dependent declines in
mitochondrial function are seen in many animals, including humans (Boffoli et al. 1994). A
recent report (Michikawa et al. 1999) shows that there are “hot spots” for age-related mutations
in the mitochondrial genome, and that mitochondria with these mutations have a higher
replication frequency than wild-type mitochondria. Thus, the mutants are able to outcompete the
wild-type mitochondria and eventually dominate the cell and its progeny. Moreover, the
mutations may not only allow more ROS to be made, but may make the mitochondrial DNA
more susceptible to ROS-mediated damage.
6. Telomere shortening:Telomeres are repeated DNA sequences at the ends of
chromosomes. They are not replicated by DNA polymerase, and they will shorten at each cell
division unless maintained by telomerase. Telomerase adds the telomere onto the
chromosome at each cell division. Most mammalian somatic tissues lack telomerase, so it has
been proposed (Salk 1982; Harley et al. 1990) that telomere shortening could be a “clock” that
eventually prohibits the cells from dividing any more. When human fibroblasts are cultured, they
can divide only a certain number of times, and their telomeres shorten. If these cells are made
to express telomerase, they can [Link], there is no correlation between telomere
length and the life span of an animal (humans have much shorter telomeres than mice), nor is
there a correlation between human telomere length and a person's [Link]-deficient
mice do not show profound aging defects, which we would expect if telomerase were the major
factor in determining the rate of aging (Rudolph et al. 1999). It has been suggested that
telomere-dependent inhibition of cell division might serve primarily as a defense against cancer
rather than as a kind of “aging clock.”
7. Genetic aging programs:Several genes have been shown to affect aging. In humans,
Hutchinson-Gilford progeria syndrome causes children to age rapidly and to die (usually of heart
failure) as early as 12 years. It is caused by a dominant mutant gene, and its symptoms include
thin skin with age spots, resorbed bone mass, hair loss, and arteriosclerosis. A similar
syndrome is caused by mutations of the klotho gene in mice (Kuro-o et al. 1997). The functions
of the products of these genes are not known,but they are thought to be involved in suppressing
the aging phenotypes. These proteins may be extremely important in determining the timing of
[Link] C. elegans, there appear to be at least two genetic pathways that affect aging.
The first pathway involves the decision to remain a larva or to continue growth. After hatching,
the C. elegans larva proceeds through four instar stages, after which it can become an adult or
(if the nematodes are overcrowded or if there is insufficient food) can enter a nonfeeding,
metabolically dormant dauer stage. It can remain a dauer larva for up to 6 months, rather than
becoming an adult that lives only a few weeks. When it comes out of the dauer stage, it will live
as long as if it had never been a dauer larva. In the dauer stage, adult development is
suppressed, and extra defenses against ROS are synthesized. If some of the genes involved in
this pathway are mutated, adult development is allowed, but the ROS defenses are still made.
The resulting adults live twice to four times as long as wild-type adults (Figure 1: Friedman and
Johnson 1988). The second pathway involves the gonads. Germ cells appear to inhibit
longevity, while the somatic cells of the gonads act to prolong the life of the nematode.
8. Changes in structural tissues:The structural integrity of the vertebrate organism depends
on two kinds of fibrous protein molecules, collagen and elastin. Collagen, which constitutes
almost one-third of the body protein, is found in skin, bone, and tendons. When first synthesized
by cells called fibroblasts, collagen is in a fragile and soluble form (tropocollagen). In time this
soluble collagen changes to a more stable, insoluble form that can persist in tissues for most of
an animal’s life. The rate of collagen synthesis is high in youth and declines throughout life, so
that the ratio of insoluble to soluble collagen increases with age. Insoluble collagen then builds
up with age as a result of synthesis exceeding removal, much like another fibrous tissue, the
crystalline lens of the eye. With increasing age, the number of cross-linkages within and
between collagen molecules increases, leading to crystallinity and rigidity, which are reflected in
a general body stiffness. There is also a decrease in the relative amount of a
mucopolysaccharide (i.e., the combination of a protein and a carbohydrate) ground substance; a
measure of this, the hexosamine–collagen ratio, has been investigated as an index of individual
differences in the rate ofaging. An important consequence of these changes is decreased
permeability of the tissues to dissolved nutrients, hormones, and antibody molecules.
The rate of aging of collagen is related to the overall metabolic activity of the animal; rats kept
on low-calorie diets have more youthful collagen than fully nourished rats of the same age.
Elastin is the molecule responsible for the elasticity of blood vessel walls. With age, progressive
loss of elasticity of vessels occurs, presumably because of fragmentation of the elastin
[Link] cross-linkage of collagen is chemically similar to the cross-linkages that occur in
skins when they are tanned to leather. This similarity has stimulated proposals that chemicals
that inhibit cross-linkage in tanning will retard aging.
9. Tissue cell loss and replacement:The tissues of the body fall into two groups, according to
whether or not there is continuous renewal of tissue cells. At one extreme are nonrenewal
tissues such as nerves and voluntary muscles, in which few new cells are formed (at least in
mammals) after a certain stage of growth. In renewal tissues such as the intestinal epithelium
and the blood, on the other hand, some cell types live only one or a few days and must be
replaced hundreds of times in the life span of even a short-lived animal such as the [Link]
these limits lie many organs, such as liver, skin, and endocrine organs, that have cells that are
replaced over periods ranging from a few weeks to several years in humans.A peripheral nerve
is a convenient object to study because the total number of fibres in the nerve trunk can be
counted. This has been done for the cervical and thoracic spinal nerve roots of the rat, the cat,
and humans. In the ventral and dorsal spinal roots of humans, the number of nerve fibres
decreases about 20 percent from age 30 to age 90. In the cat, the rat, and the mouse, however,
the data do not consistently indicate a decrease of number of spinal root fibres with age. In
humans the number of olfactory nerve fibres, which serve the sense of smell, decreases by age
90 to about 25 percent of the number present at birth, and the number of optic nerve fibres,
serving vision, decreases at a nearly comparable [Link] is a striking decrease in the
number of living cells in the cerebral cortex of the brain of humans with age. The cerebellar
cortex of the rat and human is about as susceptible to age deterioration as is the cerebral
cortex. Other parts of the brain are not so obviously marked by [Link] is, in short, a
tendency for the higher and more recently evolved levels of the nervous system to undergo
more severe aging loss than do other regions, such as the brainstem and spinal cord. It is not
yet known how much of the loss of brain cells results from conditions within the brain itself and
how much result from extrinsiccauses, such as deterioration of the blood circulation. The
nutrition and maintenance of nerve cells, or neurons, in the central nervous system depends to
a considerable extent on neuroglia, small cells that surround the neurons. The absolute number
of these cells apparently does not decrease with age, but some of the microscopic changes
seen in the neurons of old persons are similar to the changes produced by starvation or physical
[Link] has been shown that after an attack of measles, the virus remains in the host’s
body for the remainder of life and infrequently gives rise to a rapidly progressing degeneration of
the cerebral cortex. This virus or other inapparent viruses may also be responsible for the
individual differences in onset of senility in [Link] renewal tissues are typically made up of
a population of proliferative cells, which retain the capability for division, and a population of
mature cells, produced by the proliferative cells and with limited life spans. The production of
cells must balance the steady loss and also compensate quickly for unusual losses caused by
injury or disease, so each renewal tissue has one or more channels of feedback control to
adjust production to demand. Aging of renewal tissues is expressed in several ways, including
decrease in the number of proliferative cells, decrease in the rate of cell division, and decrease
in responsiveness to feedback signals. Changes of these factors in the blood-forming tissues of
the mouse are small, yet the blood-forming tissues do suffer an aging deficit, for the ability to
respond to extreme or repeated demand is significantly reduced in older [Link] intact skin
has a cell turnover time of several weeks, with the capability, shared by all renewal tissues, of
temporarily increasing the rate of cell production by a large factor in response to injury. The rate
of wound healing decreases with age, rapidly at first and more slowly as age [Link] of
the most regular and striking aging processes is the decrease in the ability to focus on both
close and distant objects. This loss in visual accommodation is the result in part of a weakening
of the ciliary muscle of the eye and of a decrease in the flexibility of the lens. A further
contributing factor, however, is that the lens continues to grow throughout life at a rate that
diminishes with age. This growth is the result of continuous division of epithelial cells near an
imaginary midline of the lens, giving rise to fresh cells that differentiate into the precisely aligned
lens fibres. Once formed, the fibres remain permanently in [Link] important feature of the
renewal mechanism is the stem cell. These cells, which may normally continue to divide at a low
rate throughout life, under conditions of increased demand enter a compensatory proliferative
phase during which they divide rapidly. Blood-forming tissue has a stem cell population that
responds to injury readily in youth, but its capacity diminishes with age. The increased incidence
of anemia in old age and the reduced capacity to respond to blood loss have been attributed to
depletion of the blood-forming stem cells. Stem cell populations have not been identified with
certaintyin other proliferative tissues. The intestinal mucosa, in particular, has a high cell-
division rate without any clear indication of a reserve population of stem cells.
 Theories on Ageing
Ageing is not just a natural loss of life but exact process of development, though vicious. A
number of factors are accountable for the ageing procedure and these jointly act through a
composite network of interacting positive and negative feedback system. The major theories
proposed to explain ageing can be generally classified into two groups. The first deals with
ageing at the level of the cell, and is known as the cellular theory, and the second takes into
account the whole body for understanding of the ageing process-the systemic theory or
extracellular theory of ageing.
Neuroendocrine Theory: All the body activities are dependent on neurological and endocrine
systems. Throughout the life, the loss of neurons and endocrine cells takes place. As a result,
there occurs reduction in brain weight due to cell loss and loss of fluids and ground substance.
Changes in endocrine functions result from cell loss and changes in receptors for hormones.
Immune Theory: The immune theory of ageing is based on two aspects: a) Age dependent
decline in the capacity of the immune system mainly thymus derived immunity (T-cell or cellular
immunity) and b) Autoimmunity – Immune system starts treating autologous tissues as foreign.
These conditions lead to vulnerability to diseases and abnormalities, consequential from
autoimmune response. Definite diseases of older age are characterized by some degree of
immune system dysfunction. These diseases comprise (i) cancer (failure of immune
surveillance), (ii) Mature-onset diabetes (autoantibody action against insulin receptors, insulin
and Islets of pancreas), (iii) Some vascular diseases and (iv) Autoimmune effects bring about
changes in proteins such as collagen and introduces errors in transcriptions, translations, or
post-translation fluctuations.
Somatic Mutation Theory: Mutations are the changes in the DNA of a cell. Somatic cell of the
body of organisms develop spontaneous mutations. These mutations get included by errors in
replication of DNA during cell division. The impaired DNA may render the cell defective in the
production of essential enzymes resulting in the cell ageing or even cell death.
Error and Fidelity Theory: The ageing results from a decline in the fidelity of all biological
components. Here Fidelity refers to true reproduction of proper proteins through transcriptions
and translation. The decrease in fidelity of transcription ortranslation is called as error and is
distinct from DNA – based mutations. Error in transcription or translation results in the addition
of improper amino acids into the sequence comprising a particular protein.
Glycation theory: The theory states that glucose acts as a mediator of ageing. The process
involves non-enzymatic reactions that take place between reducing sugars and tissue proteins
and nucleic acids. The end products of such reactions in case of proteins leads to dehydrated
and rearranged unalterable structures referred to as advanced glycation end (AGE) products.
Similar end products are produced between DNA and reducing sugars. Glycated proteins show
loss of enzyme activity, inappropriated cross linking and decreased degradation of irregular
proteins. Glycation of DNA, most usually uracil residues resulting by deamination of thymine, if
not repaired by excision repair and uracil DNA glycosylase, can induce mutation. Hemoglobin,
collagen, and lens proteins are those proteins that undergo glycation with age. Glycation of
collagen may direct to elasticity changes in blood vessels, tendons and much of the connective
tissue strain of body tissues.
Program restriction theory: It is now well recognized that the genetic information depends on
base sequences of DNA. Only small amount of the information in DNA is utilized by cells of
various tissues. Only the sequences suitable for a particular tissue are transcribed and
translated; the remaining DNA is turned off. The theory of program restriction states that ageing
may be due to increased ‘turn off’ of DNA, leading to impaired capacity of the cell to transcribe
essential RNAs including mRNAs. Though DNA segments may not be actually lost, chromatin
structure seems to change with age in a way that favours restriction of DNA transcription. This
might lead to a functional loss of portions of the gene library.
Summary:Ageing occurs in all animals and plants. It is evidenced by a decline in vital activities
of life. The symptoms of ageing are clearly demarcated at cellular, organ and system levels
including morphological, physiological and nuclear events. There is a clear relationship between
average life span and ageing of an individual’s life cycle. Numerous genetic and non-genetic
theories are proposed to describe ageing which have also been discussed in the talk.
The Free Radical Theory: Implicates the gradual accumulation of oxidative cellular damage
as a fundamental driver of cellular aging. This theory has evolved over time to emphasize
the role of free radical induced mitochondrial DNA (mtDNA) mutations and the accumulation
of mtDNA deletions. Given the proximity of mtDNA to the electron transport chain, a primary
producer of free radicals, it postulates that the mutations would promote mitochondrial
dysfunction and concomitantly increase free radical production in a positive feedback loop. It
is known that diet, lifestyle, drugs (e.g. tobacco and alcohol) and radiation etc., are all
accelerators of free radical production within the body. [4]
Error theory: based on the idea that errors can occur in the transcription of the synthesis of
DNA. These errors are perpetuated and eventually lead to systems that do not function at
the optimum level. The organism’s aging and death are attributable to these events.
The Cross-Linking Theory:[5] also referred to as the Glycosylation Theory of Aging. In this
theory it is the binding of glucose (simple sugars) to protein, (a process that occurs under
the presence of oxygen) that causes various problems. Once this binding has occurred the
protein becomes impaired and is unable to perform as efficiently. Living a longer life is going
to lead to the increased possibility of oxygen meeting glucose and protein & known cross-
linking disorders include senile cataract & appearance of tough, leathery &yellow skin.
The Neuroendocrine Theory [4][6] First proposed by Professor Vladimir Dilman and Ward
Dean MD, this theory elaborates on wear and tear by focusing on the neuroendocrine
system. This system is a complicated network of biochemicals that govern the release of
hormones which are altered by the walnut sized gland called the hypothalamus located in
the brain. The hypothalamus controls various chain-reactions to instruct other organs and
glands to release their hormones etc. The hypothalamus also responds to the body hormone
levels as a guide to the overall hormonal activity. But as we grow older the hypothalamus
loses it precision regulatory ability and the receptors which uptake individual hormones
become less sensitive to them. Accordingly, as we age the secretion of many hormones
declines and their effectiveness (compared unit to unit) is also reduced due to the receptors
down-grading
The Membrane Theory of Ageing: According to this theory it is the age-related changes of
the cell's ability to transfer chemicals, heat and electrical processes that impair it. As we
grow older the cell membrane becomes less lipid (less watery and more solid). This impedes
its efficiency to conduct normal function and in particular there is a toxic accumulation
The Decline Theory:The mitochondria are the power producing organelles found in every
cell of every organ. Their primary job is to create Adenosine Triphosphate (ATP) and they
do so in the various energy cycles that involve nutrients such as Acetyl-L-Carnitine, CoQ10
(Idebenone), NADH and some B vitamins etc. [4] Enhancement and protection of the
mitochondria is an essential part of preventing and slowing aging. Enhancement can be
achieved with the above mention nutrients, as well as ATP supplements themselves.
Cell death
Introduction to Apoptosis:Every normal living cell of animals, plants and even bacteria are
mortal. I.e., they must die after some time. Cell death is a finely tuned programme inherent in
the cells genetic machinery. This normal cell death which is the part of normal development and
maintenance of homeostasis is called apoptosis or programmed cell death (PCD).This
phenomenon is very much different from death of a cell due to pathological cause or necrosis.
This process is highly regulated and any defect in apoptotic machinery will lead to extended
survival of cells which may result in neoplastic cell expansion, leading to genetic instability
and accumulation of mutations.
Why do cells die?
Cell death is an important process in the body. It removes cells in situations including:
1. When cells are not needed, such as during certain stages of development.
2. To create a structure in the body, for ex; the outer layer of the skin is made of dead
cells.
3. To remove excess cells, such as white blood cells after an infection has been
cleared.
4. If cells are damaged, such as by radiation or toxins.
5. When cells are infected by viruses.
How do cells die?
Cells can die because they are damaged, but most cells die by killing themselves.
There are several distinct ways in which a cell can die. Some occur by an organised,
‘programmed’ process. Some cell death processes leave no trace of the dead cell, whereas
others activate the immune system with substances from the dead cell.
1. Apoptosis: is a form of cell death that prevents immune activation. Apoptotic cells have a
particular microscopic [Link] cell activates proteins called caspases that are
normally dormant. These caspases dismantle the cell from within. The apoptotic cell breaks
into small packages that can be engulfed by other cells. This prevents the cell contents
leaking out of the dying cell and allows the components to be recycled.
2. Necrosis: occurs when a cell dies due to lack of a blood supply, or due to a toxin. The cells’
contents can leak out and damage neighbouring cells, and may also trigger inflammation.
[Link]: is similar in appearance to necrosis, in that the dying cell’s contents can leak
out. However, like apoptosis, necroptosis is a programmed suicide process triggered by specific
proteins in the dying cell.

4. Pyroptosis: is a form of cell death that occurs in some cells infected with certain viruses or
bacteria. A cell dying by pyroptosis releases molecules, called cytokines that alert neighbouring
cells to the infection. This triggers inflammation, a protective response that restricts the spread
of the viruses and bacteria.

5. Cell death proteins: Many proteins have been discovered that control whether a cell dies by
the processes of apoptosis, necroptosis or pyroptosis. Some key cell death control proteins
include:

i. Caspases: these enzymes are switched on in apoptotic cells, and digest other proteins to
bring about cell death. Some caspases have roles in processes other than cell death.

ii. Bcl-2 family proteins: these proteins interact with each other to determine whether a cell
undergoes apoptosis or stays alive. Some Bcl-2 family proteins promote survival, and block
apoptosis. Others are ‘pro-death’, and trigger apoptosis.
iii. Death receptors: these are proteins on the surface of the cell. When they are bound by
certain cytokines (hormone-like signalling proteins), they cause changes in the cell that can lead
to cell death.
iv. RIP kinases: two proteins called ‘RIP1 kinase’ and ‘RIP3 kinase’ trigger necroptosis.
v. IAPs: or ‘inhibitor of apoptosis proteins’ can prevent cell death. They can do this by blocking
several cell death proteins including caspases and RIP1 kinase.
vi. SMAC/Diablo: is an inhibitor of IAPs. In healthy cells, SMAC is stored away from IAPs, in
parts of the cell called mitochondria. When cell death is triggered, SMAC can leak out and block
IAPs function. Thus, the release of SMAC out of mitochondria can promote cell death.
Process of Apoptosis :Apoptosis, the programmed cell death is characterized by chromatin
condensation and cell shrinkage in the early stage and then the nucleus and cytoplasm
fragment, forming membrane-bound apoptotic bodies which can be engulfed by phagocytes. In
contrast, cells undergo another form of cell death, necrosis, swell and rupture. The released
intracellular contents can damage surrounding cells and often cause inflammation. Apoptosis is
an important process during normal development. It also involved in aging and various diseases
such as cancer, AIDS, Alzheimer’s disease and Parkinson’s [Link] cell death, or
apoptosis, is mediated by proteolytic enzymes called caspases, which are synthesized in the
precursor forms as procaspases. When activated by various signals, caspases function to
cause cell death in most organisms, ranging from C. elegans to human beings. Apoptosis
provides a means deciding the shapes of body parts in the course of development and a means
of eliminating cells producing anti-self antibodies or infected with pathogens as well as cells
containing large amounts of damaged DNA. Cytotoxic T cells initiate apoptosis in cells to which
they bind through T-cell receptor-class I MHC-peptide interactions aided by interactions with the
coreceptor molecule CD8. Under some circumstances, such as when DNA damage is
extensive, p53 also activates expression of genes that lead to apoptosis, the process of
programmed cell death that normally occurs in specific cells during the development of
multicellular animals. In vertebrates, the p53 response evolved to induce apoptosis in the face
of extensive DNA damage, presumably to prevent the accumulation of multiple mutations that
might convert a normal cell into a cancer cell. During apoptosis, the cell is digested by a class of
proteases called caspases. More than 10 caspases have been identified. Some of them (e.g.,
caspase 8 and 10) are involved in the initiation of apoptosis, others (caspase 3, 6, and 7)
execute the death order by destroying essential proteins in the cell.
How does cell death impact health?
Many diseases are associated with Cancer cells often resist cell death, even
abnormal cell death. Some examples of after anti-cancer treatment.
this are: Cancer
Autoimmunity e.g. Lupus, type 1 Immune cells that attack the body’s own
diabetes tissues normally die. If this cell death
does not occur it can cause diseases
such as lupus or type 1 diabetes.
Viral infection Viruses need to keep a cell alive in order
to reproduce. Cell death can therefore
prevent viral replication.
Heart attack Many cells, including those in the heart
and brain, trigger their apoptosis
machinery when they lose their blood
supply.
Significance of Apoptosis
1. It helps to maintain homeostasis in the multicellular organisms.
2. Proper size of the body is maintained by apoptosis.
3. Apoptosis maintains the constancy of cell number in an organism.
4. The unwanted cells are eliminated from the body by apoptosis.
5. The dangerous T-lymphocytes are eliminated by apoptosis.
6. Programmed cell death is crucial for cell development.

Role of Apoptosis :Apoptosis plays an important role in the body of an organism. Following are
a few such roles performed by the process:
1. The separation of the fingers during the development of the foetus is due to apoptosis.
2. It results in the closure of the neural tube in the dorsal part.
3. Programmed cell death results in the removal of vestigial remnants such as pronephros.
4. During the determination of sex of the foetus, the Wolffian ducts are removed by cell death.
5. In the urachus, apoptosis allows the removal of redundant tissues between the bladder and
umbilicus.
Relationship Between Apoptosis and Cancer
Cancer is the uncontrolled division of cells that leads to the development of tumour. If the
apoptotic signalling works properly, these unwanted cells can be removed from the body. The
main reason for cancer is that they have the ability to prevent apoptosis and therefore multiply
uncontrollably.
BIOGENETIC LAW
The biogenetic law is a theory of development and evolution proposed by Ernst Haeckel in
Germany in the 1860s. It is one of several recapitulation theories, which posit that the stages of
development for an animal embryo are the same as other animals' adult stages or forms.
Commonly stated as ontogeny recapitulates phylogeny, the biogenetic law theorizes that the
stages an animal embryo undergoes during development are a chronological replay of that
species' past evolutionary forms. The biogenetic law states that each embryo's developmental
stage represents an adult form of an evolutionary ancestor. According to the law, by studying
the stages of embryological development, one is, in effect, studying the history and
diversification of life on Earth. The biogenetic law implied that researchers could study
evolutionary relationships between taxa by comparing the developmental stages of embryos for
organisms from those taxa. Furthermore, the evidence from embryology supported the theory
that all of species on Earth share a common ancestor.
Ernst Haeckel studied animals and evolution in Germany from 1834 to 1919. He proposed the
biogenetic law while working at the University of Jena in Jena, Germany, in his 1866 book
Generelle Morphologie der Organismen [General Morphology of the Organisms]. The
publication unifies theories Haeckel proposed during his work throughout the 1850s and 1860s.
Haeckel cited Johann Wolfgang von Goethe from Germany, Jean Baptiste Lamarck from
France, and Charles Darwin from England as his main influences for creating the biogenetic
[Link] proposed the biogenetic law after reading Charles Darwin's theories in The Origin
of Species. Haeckel championed Darwin's theory of evolution in Germany and praised him for
using information from embryology to help form his theory of evolution. Darwin argued that one
could explain facts about embryology, such as the early similarity between embryos of different
species, by looking at them in terms of evolution by natural selection. The fact that the more
general characters of a taxonomic group tend to be present earlier in the embryo, while
specialized and variable characters tend to manifest later in the embryo, indicated that these
specialized features are the most recent changes to the ancestral form. Darwin proposed that
the embryos of currently living species would look similar to the embryos of their ancestors and
that embryos of different taxonomic groups look similar to each other because they share a
common ancestor. Haeckel interpreted the data differently than Darwin, and he purported
instead that the embryonic stages of extant species represent adult forms of their previous
[Link] Haeckel cited Darwin as he proposed the biogenetic law, the two disagreed
about embryology and evolution.
1. Haeckel interpreted the process of evolution as progressive, following a specified path from
lower to higher animals. Darwin, however, argued that evolution wasn't progressive.
2. Darwin also argued that embryos diverged more from one another as development
progressed, rather than passing through linear stages of evolutionary ancestry.
3. Because Haeckel argued that evolution was progressive, he also endorsed Jean Baptiste
Lamarck's theory of acquired characters. Lamarck theorized that organisms could acquire or
alter their characters by use and disuse of their anatomical parts, and that parents could pass
on these acquired or altered characters to their offspring. Lamarck's theory competed with
Darwin's of natural selection as the mechanism for evolution, but Haeckel incorporated both
theories into the biogenetic [Link] proposed the biogenetic law so that researchers could
use the stages of embryological development to help construct evolutionary (phylogenetic)
trees. Haeckel claimed that phylogenesis, or the process by which groups of organisms diversify
from one another, influenced the development(ontogeny) of embryos. He theorized that the
stages in an organism's ontogeny reflected the successive changes in form, from generation to
generation, of that organism's evolutionary ancestors. Many scientists saw Haeckel's work as a
breakthrough in recapitulation theory because he offered a physical mechanism of development
that other biologists had not proposed.
Assumptions:According to Haeckel, the biogenetic law depends on three assumptions.
1. Law of correspondence:This states that each stage of development in higher animals, such
as humans, corresponds to adult stages of lower animals, such as fish. For instance, gill slits in
early human embryos correspond to the gill slits in adult fish.
2. Constraints in early development:
The second assumption of the biogenetic law was that phylogenesis must occur by the addition
of new characters to the end of the normal developmental process. Haeckel said that the early
stages of different species' embryos look similar to each other because of developmental
constraints present early in development. These constraints disappear towards the end of
development, which allow for the addition of new characters and for subsequent evolution.
3. Principle of truncation:
The third assumption was the principle of truncation. Haeckel argued that if new characters
were continuously added to the end of normal ontogeny, the length of embryonic development
would eventually become longer than gestation periods of organisms in extant species. As a
result, he theorized that early stages of development must be faster in higher organisms than in
lower [Link] also used the concept of truncation to explain inconsistencies between the
stages of animals from different taxa. For instance, pigs and humans may look similar to each
other as early embryos, but as ontogeny progresses, the embryos start to look different from
one [Link] embryos pass through the linear stages of their evolutionary ancestors, as
Haeckel claimed, then the two embryos should go through the same stages until the pig
reaches full development and the human continues through the subsequent stages of its
evolutionary ancestry. However, in many cases, scientists found no such progressions. Haeckel
hypothesized that truncation of ontogeny caused these inconsistencies. This principle of
truncation influenced scientists in the US such as Alpheus Hyatt, Alpheus Packard, and Edward
Drinker [Link] supported his biogenetic law with his drawings of embryos during
different stages of development. In 1874, his work Anthropogenie included drawings of
embryonic fish, salamanders, tortoises, chicks, pigs, cows, rabbits, and humans at different
stages of development placed next to one another for comparison. Haeckel's drawings made
the embryos of the different groups look almost identical in their earliest stages of development.
He argued that they only become recognizable as species later in their respective
developments. These similarities, according to Haeckel, demonstrated the linear progression
from what he called lower forms to higher forms of animals, and he concluded that the stages
recapitulated the evolutionary history of the organisms' ancestors. Wilhelm His, professor of
anatomy at the University of Basel in Basel, Switzerland, and at the University of Leipzig in
Leipzig, Germany, opposed Haeckel's biogenetic law. He argued that embryologists shouldn't
aim to construct phylogenetic trees and argued that embryologists shouldinstead aim to explain
development. He agreed with Haeckel that one should use causal theories to explain
development, but he argued that Haeckel's theory was flawed in positing the stages of
development as representations of adult ancestors. He argued the Haeckel's biogenetic law
overemphasized evolution as the cause of development and exaggerated the similarities
between embryos of different species. He said that there were obvious differences between the
early stages of embryos of different species, and that those differences, not the similarities,
were important to explain [Link] the decades after Haeckel's publication of the
biogenetic law, other biologists struggled to recreate Haeckel's results. Franz Keibel, a student
of Wihelm His and a professor of anatomy at the University of Strasbourg in Alsace, France,
tried to recreate Haeckel's drawings from his own specimens and concluded that Haeckel had
exaggerated the similarity between embryos in his drawings. Keibel therefore rejected the
biogenetic law and labeled it an exaggeration of the truth. In 1897, Keibel published this
conclusion in the first volume of Normentafeln zur Entwicklungsgeschichte der Wirbelthiere
(Standard Panels to the Developmental History of the Verterbra).Furthermore, many scientists
adopted a competing theory in the beginning of the twentieth century. In 1828, Karl Ernst von
Baer at University of Königsberg in Königsberg, Prussia, had proposed what researchers later
called von Baer's laws of embryology. Von Baer formulated these laws to discredit conception of
recapitulation theory published in 1811 by Johann Friedrich Meckel. In his laws, von Baer stated
that the more general characters of a taxonomic group appear earlier in an animal embryo than
the specialized characters do. He argued that rather than animals passing through successive
stages of other adult animals, they diverge from one another as development progresses.
Therefore, he concluded, the stages embryos pass through during ontogeny never represent
adult forms of other animals; they only represent embryonic stages of other animals. This
conception was part of Darwin's 1859 account of ontogeny in The Origin of Species. Although
von Baer's theory was overshadowed by recapitulation theory for most of the nineteenth
century, scientists in the twentieth century began to adopt von Baer's view as the more accurate
representation of [Link]'s biogenetic law was further discredited by the results of
experimental embryologists in the early twentieth century. Researchers abandoned Haeckel's
theory when they couldn't confirm his observations. Embryologists showed that cases of
recapitulation were less prevalent than were the inconsistencies between the developmental
stages of normal organisms from different species.
Rejections:
As per Haeckel, all animals start their embryonic development from zygote, suggesting that all
life evolved from a single cell. He believed that paleogenetic characters are ancestral traits that
are retained in the embryos and coenogenetic characters are secondary, adaptive and non-
ancestral that appear later in the development. However, Ernst Haeckel’s theory was criticised
on three counts as follows:
1) Embryos of higher animals resemble only the embryos of lower animals and not the adults,
2) Ontogeny only shows affinity and not evolution, and
3) Retaining ancestral embryonic stage has no selection value and therefore would be wasteful
and time consuming.
Modern gene theory explains the sequence of events and the subsequent end product
of ontogeny in the right perspective and, therefore, Haeckel’s original theory has now
been rejected
STEM CELLS :Stem cells are defined as cells that have clonogenic and self-renewing
capabilities and differentiate into multiple cell lineages. Stem cells are found in all of us, from the
early stages of human development to the end of life. Stem cells are basic cells of all
multicellular organisms having the potency to differentiate into wide range of adult cells. Self-
renewal and totipotency are characteristic of stem cell. Though totipotency is shown by very
early embryonic stem cells, the adult stem cells possess multipotency and differential plasticity
which can be exploited for future generation of therapeutic options. All stem cells may prove
useful for medical research, but each of the different types has both promise and limitations. For
decades, researchers have been studying the biology of stem cells to figure out how
development works and to find new ways of treating health problems. The scientific researchers
and medical doctors of today hope to make the legendary concept of regeneration into reality by
developing therapies to restore lost, damaged, or aging cells and tissues in the human body.
This research has opened new horizons for stem cell research. Stem cell research holds
tremendous promise for the development of novel therapies for many serious diseases and
injuries. While stem cell-based treatments have been established as a clinical standard of care
for some conditions, such as hematopoietic stem cell transplants for leukaemia and epithelial
stem cell-based treatments for burns and corneal disorders, the scope of potential stem cell-
based therapies has expanded in recent years due to advances in stem cell research. It is
impossible to project when actual treatments or cures might emerge from such research, but the
paths this research might take and potential applications have been much discussed. Stem cells
can now be grown and transformed into specialized cells with characteristics consistent with
cells of various tissues such as muscles or nerves through cell culture. Highly plastic adult stem
cells from a variety of sources, including umbilical cord blood and bone marrow, are routinely
used in medical therapies.
Classification of stem cells on the basis of potency
Stem cells can be classified by the extent to which they can differentiate into different cell types.
These four main classifications are totipotent, pluripotent, multipotent, or unipotent.
1. Totipotent: The ability to differentiate into all possible cell types. Examples are the zygote
formed at egg fertilization and the first few cells that result from the division of the zygote.
2. Pluripotent: The ability to differentiate into almost all cell types. Examples include embryonic
stem cells and cells that are derived from the mesoderm, endoderm, and ectoderm germ layers
that are formed in the beginning stages of embryonic stem cell differentiation.
3. Multipotent: The ability to differentiate into a closely related family of cells. Examples include
hematopoietic (adult) stem cells that can become red and white blood cells or platelets.
4. Oligopotent: The ability to differentiate into a few cells. Examples include (adult) lymphoid or
myeloid stem cells.
5. Unipotent: The ability to only produce cells of their own type, but have the property of self-
renewal required to be labelled a stem cell. Examples include (adult) muscle stem cells.
Classification of stem cells on the basis of their sources
The easiest way to categorize stem cells is by dividing them into two types: Early or embryonic
and mature or adult. Early stem cells, often called embryonic stem cells, are found in the inner
cell mass of a blastocyst after approximately five days of development. Mature stem cells are
found in specific mature body tissues as well as the umbilical cord and placenta after birth.
1. Embryonic stem cells: Embryonic stem cells are self-replicating pluripotent cells that are
potentially immortal. They are derived from embryos at a developmental stage before the time
of implantation would normally occur in the uterus. The embryos from which human embryonic
stem cells are derived are typically four or five days old and are a hollow microscopic ball of
cells called the blastocyst.
2. Adult stem cells: Adult stem cells are undifferentiated totipotent or multipotent cells, found
throughout the body after embryonic development that multiply by cell division to replenish dying
cells and regenerate damaged tissues. The primary roles of adult stem cells in a living organism
are to maintain and repair the tissue in which they are found. Unlike embryonic stem cells,
which are defined by their origin (the inner cell mass of the blastocyst), the origin of adult stem
cells in some mature tissues is still under investigation.
3. Pluripotent stem cells I: Recently, a third type of stem cell, with properties similar to
embryonic stem cells, has emerged. Scientists have engineered these induced pluripotent stem
cellsi (iPS cells) by manipulating the expression of certain genes - 'reprogramming' somatic cells
back to a pluripotent state.
 METAMORPHOSIS
The phenomenon of metamorphosis is defined as a 'process during development which involve
a dramatic change in morphology and physiology of the larva,so that it is transformed into an
adult with completely different morphology and physiology, often for life in a different [Link]
many such anfrom the adult. The best known examples are tadpoles of frogs, caterpillar of
butterflies and moths, tadpole larva of ascidians, various larval types of crustaceans, ciliated
uochophores of marine annelids and molluscs etc;In fact sometimes the difference between the
larva and adult is so great that without knowing the origin of the egg, or without following the
young one through its full development, it would be next to impossible to know that the young
and the adult are of the same [Link] the past such differences have sometimes led to the
larva and adult of the same species being assigned to different taxonomic groups. For example
the larva and adult of the axolotl Ambystoma (Urodele--amphibia) and Tribegulians of blister
beetle (Inaecta) till quite recently were mistakenly assigned to different species. &re than a
century ago, only the study of metamorphosis of the tadpole larva of ascidians could decide that
the ascidians belong to Phylum [Link] changes that occur due to metamorphosis relate
often to a change in habitat with a corresponding change in the organisms' suucture and other
[Link] example,in sea urchins, there occurs a change from planktonic to benthic
existence,in frogs and toads from an aquatic to a terrestrial m'bde of life and in insects from a
non-flying to a flying [Link] the present unit, which deals with metamorphosis, you will be
studying this phenomenon in two diverse animal groups: namely amphibians among the
vertebrates and insects among the [Link] study will show that the metamorphic
process in animals differ in the nature of transformation and in the mode of causation, making it
impossible to describe metamorphosis in a generalized manner; however it must be pointed out
that there are certain basic [Link] all these animals, development does not stop at
hatching but continues during post embryonic life, and the principles that apply to embryonic
development also apply to post embryonic development. Furthermore, the post development
process in such animals are usually found to be reactivated by specific hormones, which modify
& modulate the timing & duration of the [Link] metamorphic changes involve differential
destruction of certain tissues accompanied by increase in growth & differentiation of other tissue
TYPES OF DEVELOPMENT
Direct development:In some animals whose eggs have little or no yolk, such as the placental
mammals,the embryos develop and grow within the womb of the mother from which they also
obtain nourishment. Once, the offspring has developed to a stage where it looks like a miniature
adult, the mother gives binh to it. This young one achieves its adult size & sexual maturity after
birth over a period of time by gradual [Link] animals whose eggs have a large amount of
yolk. such as those of the birds and reptiles, the eggs develop oviparously outside the mother's
body. Young ones resembling the miniature adults hatch out. The young individual then attains
adult size and sexual maturity by gradual growth over a length of time.
Indirect development:In many animals like frog. Amphioxus, Herdmania, insects and many
other invertebrates and vertebrates, the offspring which hatches out from the egg looks very
different from the adult form, and is called a larva. The larva leads an independent existence for
some time that varies from species to species, and then transforms into a miniature adult by the
process of metamorphosis.
TYPES OF METAMORPHIC CHANGES
The process of metamorphosis involves reactivation of the morphogenetic processes. The
morphogenetic changes as well as the mode of causation of these changes vary in different
animal groups. The degree of changes that occur during metamorphosis depend on the degree
of difference between the larva and the adult forms. For ex, in urodele amphibians (newts) and
hemimetabolous insects (cockroach), larva and adult show a few [Link] metamorphic
changes are relatively less and metamorphosis is said to be gradual or incomplete. On the other
hand, in anuran amphibians (frogs) and holometabolous insects, where differences between the
larva and adult are enormous, the changes during metamorphosis are extensive and drastic.
This type of metamorphosis is called radical or complete [Link] changes during
metamorphosis, include those of structural, physiological and biochemical nature. These are
marked by disintegration and atrophy of some structures, cellular death in some tissues,
morphogenesis and differentiation of certain new structures and remodelling of some others.
These changes of metamorphosis in at least some groups of animals (insects, crustaceans,
amphibians), are known to be controlled by hormones which serve as the causative agents of
metamorphosis.
METAMORPHOSIS IN AMPHIBIANS
Metamorphosis is radical in anurans, slight or absent in urodeles. In anuran amphibians like
toads and most frogs, metamorphosis is usually associated with a transition from an aquatic to a
terrestrial or amphibious mode of life. Occasionally however no transition in mode of life occurs
as observed in the larval and adult of the frog Xenopus laevis and many primitive anurans which
remain aquatic throughout their life. The change in habitat in the frogs and toads also usually
results in a change in their feeding habit. In some like Xlaevis there is no change in food habit
since both larvae and adult ale [Link] anurans undergo an abbreviated type of
metamorphosis before hatching, as they pass through a tailed, gilled tadpole-like stage within
the jelly membrane of the [Link] undergo direct development by skipping the larval stages
[Link] in urodele amphibians is usually less striking. Some of them undergo
direct development, while others fail to complete their [Link]. latter achieve sexual
maturity as larvae, as seen in the axolotl larvae of Ambystoma. This phenomenon is called
[Link] urodeles like salamanders have been observed to undergo two metamorphosis.
Metamorphosis in both anurans and urodeles essentially involves the activation of the genomic
set underlying the adult organization, which requires for its expression a minimum mass of
tissue that is greater than that of the [Link] activation is believed to be due to the secretion .of
a brain hormone which initiates metamorphosis. The hormone triggers the degeneration of
redundant larval organs and growth of hitherto quiescent structures which are needed in the
[Link] amphibians the process of destruction and growth are smoothly coordinated, as a
result of which the animal retains its functional integrity throughout metamorphosis instead of
lying dormant as in the case of insects.
The process of metamorphosis in anurans:Metamorphosis is most dramatic in the anurans.
In them it represents an intensive period of growth and developmental changes during which the
suucture and function of almost every organ in the body is radically modified for adaptation to a
new way of life. Unlike other adaptational changes, however, this process is begun and normally
completed in the anticipation of the change of the environment. Some changes seen during this
process involve growth and maturation of the tissues [Link] changes result in the
regression and death of organs; the gills and tails which become redundant in adult life are
completely [Link] most familiar kinds of anurans are frogs whose eggs develop into
Ladpoles, which are limbless, aquatic creatures with an oval torso and long, finned.
Metamorphosis transforms these tadpoles into four-legged, hopping carnivorous [Link]
metamorphosis most types of anurans lead a terreslrial existence. returning to the water only to
[Link] also exhibit this type of life [Link] have many anatomical and physiological
adaptations that distinguish them from the [Link] fish like tadpole is herbivorous, as a result
of which its mouth has two horny beaks with rows of horny teeth which help in rasping away
plant tissues. On each side of the head a fold of skin, the operculum, is present which covers
the gill that grow from the lower ends of the visceral [Link] are rudimentary and non
[Link] epidermis has large pigment cells of different kinds and is, underlaid by a jelly-
like dermis rich in hyaluronic acid. Because of its herbivorous diet the intestine of the tadpole is
long and [Link] eyes are r&ssed in the head, and the visual pigment porphyropsin is
present in the retina. Nitrogenous waste products are excreted primarily as ammonia. The red
blood cells are produced mainly in the kidney and the haemoglobin (HbF) present in the
tadpoles is different from that of adult (HbA).In order to transform into adults these tadpoles
undergo three distinct phases of metamorphic changes which are visible morphologically, and
have been described by Etkin as: (i) Premetamorphosis is defined as an initial period of
extensive growth but little developmental change. This is followed by (ii) Prometamorphosis
during which growth continues but conspicuous developmental changes such as the growth of
hind legs also take [Link] ends with the emergence of the Forelimbs and
then the third and final stage of development, (iii) the Metamorphic climax sets in. This is a
comparative brief period in which profound morphological changes occur very rapidly, the most
conspicuous being the complete resorption of the large muscular [Link] tadpole is transformed
into an immature froglet and metamorphosis is [Link] length of the larval period in
anurans vary. In some species, tadpoles produced in spring metamorphose during early
summer. while in others the tadpole stage may last for a year or more before the
premetamorphic transformation [Link] first sign of change occurs in the growth or
premetamorphic period by the appearance of swellings on each side at the posterior end of the
[Link] are the hind limbs which grow slowly during the prometamorphic [Link] growth
period is followed by the prometamorphic period during which the lengt of hindlimb increases
very rapidly relative to the growth of torso.A few days before the end of the prometamorphic
period a number of changes begin, notably a shift in the position of the anus and a thinning of
the operculum on each side, a process that forms translucent, "skin windows" through which the
forelimbs will subsequently eruptThis is followed by metamorphic climax, as a result of which
the the forelimbs grow and erupt through the operculum, the horsy beaks of the oral region are
lost, the mouth widens and muscular jaws develop. The eyes are repositioned to a higher level
and develop [Link] changes involve a complete remodelling of the head skeleton and are
adaptive to a predatory mode of life, requiring an aerial sensory input. The epidermis thickens,
hardening the skin and the jelly-like dermis gets replaced by a tougher more librous tissue.
Within the skin, the pigment cells get arranged in such a manner so as to give the adult pattern
of colouration. A muscular tongue used for capturing prey develops, the hyoid cartilages
differentiate, the gills are resorbed, as lungs for pumping air into the body which become fully
[Link] cells of the larval alimentary tract are almost completely sloughed off and
essentially a new and shorter digestive tract develops. Within a week of metamorphic climax the
tadpole transforms from a tadpole to a [Link] melamorphosis some anurans adopt a
terrestrial mode of life,returning to the water only lo breed. Others spend a considerable amount
of time out of water but usually remain in a moist environment.
i) Morphological changes:The changes in the structure of amphibians during metamorphosis
are grouped into there categories and are described as a) Regressive changes-These
changes involve the gradual reduction and ultimate disappearance of all those larval structures
or organs which become redundant in [Link] ventral suckers, external gills, the long tail with
fin folds are reabsorbed during early functional life. The gill clafts are closed; the peribranchial
cavities, the horny teeth and horny lining of the jaws are [Link] shape of the mouth changes,
the cloaca1 tube shortens and gets reduced. The lateral line organs of the skin of tadpole
disappear and some blood vessels are reduced b) Progressive changes-Some organs and
structures become functional during and after metamorphosis and involve the development of
the fore and the hind limbs,the middle ear in connection with the first pharyngeal pouch (the
pouch situated between the mandibular and hyoid arches), the tympanic membrane supported
by the circular tympanic cartilage. The eye protrudes on the dorsal surface of the head and
develops an upper eyelid. The tongue develops from the floor of the mouth c) Remodelling -
Some structures and organs which occur and function both before and after metamorphoses,
get transformed or remodelled during the process in order to meet the requirement of the adult
mode of [Link]
changes affect primarily skin, intestine and [Link] skin thickens, and becomes glandular by
[Link] mucous and serous glands. It also develops an outer keratinized layer
as well as characteristic colour and pattern of pigmentation. The brain gets highly differentiated.
The intestine which was long and coiled in the herbivorous tadpole shortens and straighten out:
Other notable changes which occur are the change in the blood vascular system in order to
supply the lungs, the change in the portal system, the change in the heart as it becomes three
chambered from being two chambered earlier.
(ii) Biochemical changes:Biochemical changes also accompany morphological changes and
are quite striking in anuran metamorphosis.a) The porphyropsin (a complex between the protein
opsin and aldehyde of Vitamin A2) of the eye during metamorphic climax is replaced almost
completely by rhodopsin (a complex between the protein opsin and aldehyde of vitamin Al) as
the visual pigment. In the eye the larval form of a crystallin in the lens is replaced by an adult a
crystallin that differs markedly in electrophoretic mobility. An adult form of skin keratin replaces
the larval form at metamorphosis. During metamorphosis large quantities of hyaluronidase are
produced. This enzyme eliminates the hyaluronic acid of the larval skin. This hyaluronic acid is
almost replaced by a mixture of other glycosamino [Link] is not present in
appreciable quantities in the adult [Link] biochemical changes that occur in the skin as a
result of metamorphosis are alteration in the patterns of collagen synthesis and deposition. This
results in a tougher skin, more appropriate for a land dwelling existence.b) After metamorphosis,
the frog begins to excrete the bulk of its nitrogenous waste as urea rather than ammonia. Urea
like ammonia is very soluble in water but less toxic. As a result after metamorphosis urea can
be formed and retained in the blood and then be excreted by the kidneys with less water loss
than would be required for elimination of an equivalent quanity of nitrogen in ammonia form.
Production of urea requires the activation of the ornithine-urea cycle in the [Link] this cycle
carbon dioxide and nitrogen are excreted in the form of urea. Metamorphic changes thus involve
a reorganization of the metabolic patterns in the liver for production of the appropriate enzymes
of the ornithineurea cycle. This reorganization starts very early in metamorphosis even before
there is any apparent change in the body form.c) At metamorphosis the site of erythropoiesis
shifts from the liver to the bone marrow and spleen. This shift is marked by the production of
haemoglobin with different physiological and electrophoretic properties. In Rana catesbeiana
the shift from production of tadpole haemoglobin (HbF) to adult haemoglobin(HbA) is essentially
complete just before tail is resorbed. Also during metamorphosis there is considerable synthesis
of hydrolytic enzymes involved in the resorption of larval gut and tail.d) Degrowth occurs when
feeding by the larva is suspended during [Link] larva at this stages utilizes the
reserve food to produce the energy needed for completing the various metamorphic processes.
As a result there is loss of weight, with the consequence that the miniature frog produced at the
end of the metamorphosis is smaller & lighter than the mature [Link] reduction of body mass
is termed as 'degrowth'. The reduction in body mass is also partly due to shrinking of some body
parts of the larva. For instance during metamorphosis the head and trunk become smaller and
some parts like the tail and gills are loste) Autolysis causes the disappearance of larval tail, gills,
external gills and fin [Link] process of autolysis is brought about by active movement of
amoeboid macrophages which phagocytose the debris of disintegrating [Link] lysosomal
enzymes of the phagocytes, in particular the cathepsin, show a high rise in concentration and
[Link] phagocytosed or autolysed material is reabsorbed and is used in the construction
of new organs of adult. f) Enzymes during metamorphosis show a remarkable change. There is
a change both in the types and amount of enzymes produced to help in the completion of
metamorphosis. Some larval enzymes not needed in adult are no longer synthesized, whereas
new enzymes needed in the adult begin to be [Link] of carbohydrates, lipid
and nitrogen undergo changes in the adult
The process of metamorphosis in urodeles:Metamorphosis in urodele amphibians is
considerably less [Link] larval salamander (a member of genus Ambystoma) for instance
has external gills and a long tail with dorsal and ventral [Link] broad mouth, both of the larva
and the adult remains essentially the same. The forelimbs appear earlier than the hind limbs
and both pairs of appendages grow gradually, independent of any metamorphic stimulus.
Metamorphosis essentially involves loss of external gills and tail fin, development of lungs,
closure of gill slits, appearance of eyelids, formation of maxillary bones,ossification of skeleton,
cornification of skin and differentiation of skin glands. In urodeles, metamorphosis is more
gradual on the whole and may take several weeks. It involves the following changes:
a) Regressive changes: Tail is retained but fin fold disappears. Branchial apparatus gets
reduced. The external gills get reabsorbed and gill clefts close. The visceral skeleton
becomes reduced.
b) Progressive changes:Head shape changes. The eyes develop lid and bulge more on the
dorsal sides of the head. Skin becomes multilayered and cornified. Its pigmentation changes
and the skin glands also become differentiated. The legs and alimentary canal hardly undergo
any [Link] metamorphosis, depending on the species, urodeles may reduce the time
spent in water or they may become land dwellers returning to the water only to breed. Newts
and Salamanders that become terrestrial undergo a second metamorphosis when they return to
the water for [Link] metamorphosis-Some urodeles like newts and salamanders
show two major changes in form, physiology and life habit between birth (or hatching) and
[Link] example, the spotted newt Notophthalmus viridescens, exhibits two prominent
metamorphic changes during its lifecycle. This newt hatches out as an olive green larva with
gills, a keeled tail and a well developed lateral line organ system (a system of receptors
sensitive to local deplacement of water). After a few months of growth it undergoes
metamorphosis, whereby the gills and tail fins are resorbed, the lateral line system becomes
nonfunctional and skin becomes rough and dry and orange in [Link] visual pigment which
was mainly porphyropsin in the larva changes to a mixture of porphyropsin and rhodopsin. The
newt is called an eft and it becomes a woodland dweller for 2 or 3 years, remaining on land and
growing to full [Link] eft phase is terminated when the animal undergoes a second
metamorphosis through the action of hormones prolactin and pituitary gonadotropins. These
initiate a drive towards water which is accompanied by such physiological changes as the
functional reinstatement of the lateral line organ system, the reversion of the skin to a mucous
secreting, wet and shiny organ, the restoration of a finned tail, the maturation of the gonads and
the adoption of porphyropsin as the main visual pigment. In this manner the newt returns to the
water, to spawn and remains there throughout its life which lasts for several breeding seasons.
Some urodeles, like the permanently-gilledsalamanrlcrsa ppropriately called 'perinnibranchiate',
do not undergo metamorphosis in nature and so grow to sexual maturity as permanently aquatic
animals that retain their larval features like external [Link] they are neotenic and
[Link] is a condition in which the larval characters are retained for prolonged
periods of [Link] is the process by which larval individuals reproduce.
Hormones in metamorphosis of amphibian:In amphibians, the changes that occur during
metamorphosis are brought about by hormonal secretions of the thyroid gland. The first
indication of this was obtained in 1912 when Gudematsch reported that frog tadpoles when fed
dried and powdered sheep thyroid underwent precocious or early metamorphosis. Similar
results however were not observed when tadpoles were fed with preparation of other glands.
These experiments made it possible to postulate that thyroid hormones bring about
metamorphosis. In 1918 B.M. Allen observed that removal of thyroid rudiment from early frog
tadpoles, prevented them from undergoing metamorphosis, and caused them to become giant
tadpoles instead. On the other hand, if they were fed with thyroid or immersed in water
containing soluble extracts from thyroid glands they then proceeded immediately to
[Link] experiments in urodele amphibians, in particular in axolotl, Ambystoma
mexicanum, also indicated the importance of the thyroid gland in urodele metamorphosis. Later
studies by W. Etkin 1968 have also shown the importance and role of different hormones in
metamorphosis. He concluded that development is controlled by a dynamic balance of plus and
minus factors or hormones. Furthermore he found that the spacing of metamorphic events
depends on the concentration of thyroid hormone,while the sequence of events is inherent in
the tissues.
Metamorphosis in frog
‘It is a biological process by which an animal physically develops after birth or hatching,
involving a prominent change in the animal’s body structure through cell growth and
differentiation’. In other words, ‘Metamorphosis is a post-embryonic extension of the
developmental potential and involves dramatic changes in habit, habitat, morphology,physiology
and behaviour of larva so that it is transformed into the adult having entirely different habitat and
structure’. To compensate deficiency of yolk in egg, frog develops indirectly through an
intermediate free living stage, Tadpole.
Tadpole features
Oral sucker stage: • One pair of oral suckers on ventral side of head are the only distinct part
which help larva attach to any object for a couple of days •No limb buds or tail or other body
parts are distinct.• Does not feed or move
External gill stage:The buds of hind limbs come out, and tail starts developing with dorsal and
ventral [Link] pairs of external gills come out on lateral sides called [Link] eyes
become [Link] jaws develop with teeth for herbivorous mode of [Link] to 40-
50mm size after swimming about for 3-4 weeks at 26-28 degree C temperature.

Internal gill stage:Fish-like appearance with well developed internal gills covered with
operculum and have [Link]-like two chambered venous [Link] ammonotelic
(excretes ammonia as wastes), nephrostomes connected to [Link] developed tail with
dorsal and ventral [Link] developed hind [Link] and lateral line system well developed.
Alimentary canal elongated for herbivorous mode of feeding and digestion.
Changes during metamorphosis from tadpole to adult:
1. Retrogressive changes: Disappearance of tail. Loss of gills, gill chambers and
operculum. Loss of lateral line system. Lungs are formed Horny jaws replaced by bony
structures

[Link] changes: Hind limb and pelvic girdle fully formed. Fore limbs are formed
below the opercula. Eyes become enlarged, protrucible and nictitating membrane is formed.
Maxillary and Vomarine teeth are fully formed on the upper jaw. Middle ear and tympanum is
fully formed.

3. Physiological changes Shortening of alimentary canal occurs due to change in

herbivorous to carnivorous habit. Fish-like heart changes to three chambered heart
and plan of circulatory system changes due to change in respiratory system accordingly.
Nephrostomes get disconnected from nephrones, and mode of excretion changes from
ammonotelic to ureotelic. Liver function changes due to change in feeding habit.
Increase in carbohydrate metabolism occurs. Endocrine function of pancreas begins,
insulin is secreted. Visual pigments of tadpoles are porphyropsin (retinene 2), while
during metamorphosis there is a shift to the use of rhodopsin (retinene 1). The
reduction of the gills and tail is affected by autolysis of the component tissues of these
organs, with active participation of amoeboid macrophages, which phagocy tose the
debris of the disintegrating cells Biochemical metamorphic alterations may be
considered to have direct adaptive value or to serve as a basis for morphological,
chemical or other changes which have adaptive value relating to the transition from
water to land. Shift from ammonotelism to ureotelism, increase in serum albumin and
proteins, alterations in the properties and biosynthesis of haemoglobin are the
important adaptive changes. Development of digestive enzymes also contributes to the
success of the differentiation. Major modifications occur in water balance, visual
pigments, pigmentation, and tail metabolism, which aid in adjustment to land.
Hormonal Control of Amphibian Metamorphosis: During metamorphosis concurrent
changes in all body parts suggest the existence of hormones released in large quantities from the
thyroid gland of the animal. This indication was given by Gundernatsch (1912) when he fed
some frog tadpoles on dried and powdered sheep thyroid gland and observed their
metamorphosis precociously

Role of prolactin hormone:Another anterior pituitary hormone, called prolactin is also found
to be involved as an inhibitor in the overall control of metamorphosis. Developmental control
is effected by a balance between inhibition and stimulation at the level of endocrine action.
Thyroid hormones are also known to affect the process of protein synthesis at the levels of
transcription and translation and to have a role in cell differentiation ,growth and development.

Neuroendocrine control of metamorphosis:The amphibian metamorphosis is under

neuroendocrine control, involving neurosecretory cells in the brain (the hypothalamus) and two
endocrine glands, the pituitary (anterior pituitary)and the [Link] trigger to metamorphosis
may be an environmental signal affecting the larval brain through the nervous system, or there
may be an endogenous ‘clock’ in the [Link] a way, hypothalamus integrates

theinformation received from body with the environmental information  .

Function of Metamorphosis:Scientists remain uncertain why metamorphosis evolved. For
the animals of today, its purpose is obvious: if metamorphosis did not occur, tadpoles could not
become frogs and larvae could not become full-grown adults capable of reproduction. Without
reproductively mature members, these species would quickly die [Link] why would these
species evolve to need this extra step in the first place? Why not just hatch full-grown butterflies
or frogs from eggs?At least some metamorphosing species did not start out that way: the
earliest insects basically did hatch as full-grown adults. But a few hundred million years ago,
some species stumbled upon the trick of metamorphosis. It was apparently wildly successful; it
is thought that almost two-thirds of species alive today use metamorphosis to accomplish large
changes between their adult and juvenile forms .The benefit of metamorphosis may lie in its
ability to reduce competition. Pre-metamorphic animals typically consume completely different
resources from their adult [Link] live in water, eating algae and plants. Frogs live on
land, breathing air and eating [Link] eat leaves; butterflies live off of nectar. Etc..
This effectively prevents older members of the species from competing with younger members.
This may lead more members of the species to successfully reach sexual maturity, without the
risk of being out-competed by older members of their species.
Types of Metamorphosis
Complete Metamorphosis:In complete metamorphosis, a larva completely changes its body
plan to become an [Link] most famous example is that of the butterfly, which starts out as a
worm-like, leaf-eating caterpillar and transforms into a flying, nectar-drinking creature with an
[Link] that undergo complete metamorphosis are called “holometabolous,”
from the Greek words “holo” for “complete” or “whole,” “meta” for “change,” and the noun “bole”
for “to throw.” “Holometabolous,” then, means “completely changing,” or “wholly changing.” This
transformation is so swift and complete that the caterpillar must spin a cocoon and lie dormant
for weeks while its body undergoes these radical changes. Other animals which transform from
a worm-like larval stage into an animal that looks completely different include beetles, flies,
moths, ants, and [Link] scientists believe that the larval stage of complete metamorphosis
may have evolved from insects which hatched from their eggs without developing properly.
Some of these embryos may have survived long enough to find food in the outside world; and
this may have ended up giving them an advantage, as they would be able to feed longer & gain
more strength than their peers before metamorphosing into the adult stage.
Incomplete Metamorphosis:In this, only some parts of animal’s body change during
metamorphosis. Animals that only partially change their bodies as they mature are called
“hemimetabolous,” from the Greek words “hemi” for “half,” “meta,” for “change,” and the verb
“bole” for “to throw.” “Hemimetabolous,” then, is a word meaning half-changing. Cockroaches,
grasshoppers, and dragonflies, for example, hatch from eggs looking a lot like their adult selves.
They do acquire wings and functioning reproductive organs as they grow, but they do not
completely remake their bodies like their completely metamorphosing cousins do.
Examples of Metamorphosis:Frogs:The metamorphosis of a tadpole into a frog is a little less
violent than that of a caterpillar into a butterfly, but the processes share some important
common features. Tadpoles do not dissolve their bodies into mush; but they do “digest” them in
a less spectacular way. Using the process of apoptosis – or “programmed cell death” – the
tadpoles “order” the cells they don’t need anymore to shred their DNA and die. The dead cells
are then cannibalized for energy and raw materials to make other [Link] cells of their tails are
broken down and used to make their developing legs; a similar process happens with the gills,
which disappear as the tadpole begins to develop air-breathing lungs. One interesting thing to
note is that tadpole metamorphosis and insect metamorphosis likely developed separately; the
common ancestor of insects and amphibians diverged long ago, and the ancestors of modern
insects are not thought to have used metamorphosis. When the same phenomenon evolves
twice in radically different organisms, that’s a sure sign that it is a useful adaptation.
Thyroid gland regulation in amphibian metamorphosis involves the hypothalamus, pituitary
gland, and thyroid gland itself, forming the hypothalamo-pituitary-thyroid (HPT) axis. The
hypothalamus releases corticotropin-releasing factor (CRF), which signals the pituitary to
secrete thyroid-stimulating hormone (TSH). TSH then stimulates the mature thyroid gland to
produce thyroid hormones (TH), primarily thyroxine (T4) and triiodothyronine (T3). These
hormones act on target tissues, initiating the complex cascade of gene expression that
underlies amphibian development and tissue remodeling.

The HPT Axis and Hormone Production:

1. Hypothalamus: Signals the pituitary by releasing CRF.

2. Pituitary: Responds to CRF by secreting TSH.
3. Thyroid Gland: Stimulated by TSH, it matures and increases its secretion of T4 and T3
hormones.

Hormone Action and Gene Regulation:

1. Thyroid Hormones (TH): T3 is generally considered the more potent hormone.

2. Thyroid Hormone Receptors (TR): In the nucleus of target cells, TRs, when bound by T3,
activate or repress gene transcription, leading to metamorphic changes.
3. Gene Expression: TH-mediated changes in gene expression control the development of
adult organs (like the limb and intestine) and the regression of larval structures (like the tail
and gills)
 METAMORPHOSIS IN INSECTS
General process of post-hatching growth in [Link] a young insect hatches out from an
egg it is covered by a firm, inflexible,sclerotized cuticle, which due to its rigid structure cannot
grow along with the increase in length of the hatched larva. Thus during development, for the
hatched larva to attain its adult size and form, the old rigid cuticle at every stage of growth has
to be replaced by a newer, larger one by the process of moulting. Accordingly development is
marked off by a series of [Link] interval between the two moults is known as stadium and
the form that an insect assumes as a result of the moults is called instar. All insects undergo
several moults after hatching from the egg and before emerging as adults which are called
[Link] number of moults is 4 or 5 in an insect species and is usually fixed or predetermined,
but it is not absolutely constant. Furthermore, the form of the insect changes with each moult in
a precise pattern characteristic of the species. Moulting is not just a mechanical process;
instead at each moult, the moulted insect undergoes changes both in its cuticular covering and
internal organization. So you can see that development in insects is usually by the process of
metamorphosis. The metamorphic changes may be siight and gradual or radical as occurring
during the pupal [Link] this stage the lama remains quiescent without feeding or
movement, while many larval characters disintegrate and adult structures are formed. Pupal
stage follows the last larval molt and from the pupa the imago emerges
Patterns of metamorphosis
(1) Slight or no metamorphosis as observed in Apterygota-ametabolous or direct development.
(2) Incomplete metamorphosis as observed in Exopterygola-Hemimetabolous development.
(3) Complete metamorphosis as observed in Endopterygola-Holometabolous Development
(1) Slight or no metamorphosis:In primitive wingless insects (Apterygota) like spring tails,
silver fish etc. and in secondarily apterous insects the young ones that hatch from the egg are
essentially similar to imago, differing from it only in size, immature reproductive organs and
external genitalia. It moults several times leading to a gradual increase in size accompanied by
the appearance of the external genitalia and maturation of [Link] apterygotes, growth
and moulting are seen to continue even after reproductive [Link] primitive insects
undergo little or no true metamorphosis during their life cycles and these insects referred as
ametabolous. For example: Silver fish, firebrats and [Link] ametabolous insects,
immature emerges from the egg looks like a tiny version of [Link] only difference between
adult stages & juvenile stages is that in juvenile stages the genitalia are not mature & juveniles
are smaller than the adults.
Egg -> Young goes through several stadia and moults -> Imago (Adult)

(2) Incomplete metamorphosis:In the Exopterygote insects rudiments of wings are present
only as buds at hatching and the body form is disproportionate to that of the adult. As the moults
occur the form and size of the larva progressively approach that of the adult. The wings become
fully developed and sexual maturity is achieved at the last moult. The degree of metamorphosis
in the hemimetabolous forms are of two distinct types, namely gradual metamorphosis and
extensive [Link] this, three life stages occur-egg, nymph and [Link] type of
transformation refers as incomplete metamorphosis. For example: grasshoppers, mantids,
cockroaches, termites, dragonflies and [Link] hatch from their eggs look like miniature
adults. These young insects are called [Link] each shedding of skin, nymph enters a new
“instar”, a new stage of [Link] resembles the adult but lacks wings and genitalia .
(a) Gradual metamorphosis - In Exopterygote insects like grasshoppers, crickets,cockroaches
etc., the nymph is similar in both structure as well as in habit to the adult, but lacks wings,
gonads and external genitalia. It undergoes several moults as it grows and develops genitalia,
gonads and wing pads in the later larval period. The wing pads get transformed into functional
wings at the last moult, after which no more moults occur. This type of metamorphosis has no
quiescent stage and there is no loss or remodelling of larval parts.
Egg -> Nymph ->several instars and moults---> Imago (Adult)
(b) Extensive remodelling - In insects like damsel flies, mayflies, dragonflies etc.. the nymph
differs considerably in both external structure as well as in habits from the adult. The nymphs
are aquatic and herbivorous and possess some organs for locomotion in water, tracheal gills for
respiration and mandibulate mouth parts for cutting vegetation. Such aquatic larval forms are
called naiads. They undergo extensive remodelling to assume the adult form. They shed gills
and modify their mouth parts and develop wings. Moulting may lead to one or more quiescent or
semiquiescent larval stages. Egg -> Naiad -> several moults -> Imago (Adult)
(3) Complete Metamorphosis:In all Endopterygote insects, in which wings and other
structures develop internally, (in invaginated imaginal epidermal pockets) like beetles, wasps,
bees, butterflies moths etc., the larva which hatches out of egg is very different from the
imago,habit, appearance and structure. The larva has a worm-like body, biting and chewing
mouth parts, simple eyes and weakly developed walking legs. It has quite a differenthabit. For
example the mosquito larva lives in water and feeds on protozoa and algae,while the adult
either sucks blood or fruit and flower juices. A more common example is the butterfly larva
which crawls on the ground and feed on leaves and then gets transformed into an aerial
organism feeding on nectar from [Link] larvae of these types of insects are either
swimmers or crawlers, and are voracious eaters. They grow in size and moult several times till
they attain a quiescent, nonfeeding stage called pupa. The pupa is enclosed in a pupal case or
the puparium secreted by the labial glands of the larva. This pupa does not move or feed and its
energy must come from the nutrients it ingested while a larva. Externally it appears as an
inactive structure. However internally it undergoes basically two types of changes at a rapid
pace and moults only after the complex series of internal changes have taken place. These
changes involve wholesale destruction of most of the larval tissues (histolysis) and the
formation of an entirely new adult body whose organs and systems are developed
(histogenesis) from nests of organ specific cells, called the imaginal discs. Histolysis results in
systematic destruction of the old body of the larva so that all the organs except for the central
nervous system are broken down by special amoebocytic cells called phagocytes. The tissue
fluid, which arises due to the destruction is used as raw material in +e formation and
histogenesis of the adult organs. After these changes are completed as a result of histolysis and
histogenesis the pupa undergoes the pupal moult and the imago (adult) emerges fully ready to
lead a short or long independent existence and to reproduce. In this, four stages in life cycle
occur-egg, larva, pupa and [Link] is characterised by a radical change in form and
ecological habits between immature and [Link] ex: butterflies, moths, flies, ants, bees and
[Link] of the old body of larva is systematically destroyed by apoptosis, while new adult
structures such as legs, wings, structure on head, genitalia and part of changing epidermis
develop from undifferentiated nests of [Link], within any larva, there are two distinct
populations of cells: i) Larval cells- used for functions of juvenile insect. ii) Thousands of
imaginal cells- lie within larva in clusters, awaiting the signal to differentiate. Larva (caterpillar,
grub, and maggot) undergoes a series of molts as it become [Link] between these larval
molts are called [Link] does not feed and is usually considered as resting [Link]
the pupal stage, the body undergoes a complete reorganisation, transforming into the adult.
Egg -> Larva -> several instars and moults -> Pupa -> pupal moult->Adult.
Imaginal Discs: In the holometabolous larva, there are two cell populations: (1) the larval cells
which are used for the larval structures and (2) the imaginal disc and the histoblasts which are
present in clusters, awaiting the signal to differentiate. Imaginal discs do not occur in the
nymphs and larvae of hemimetabolous [Link] imaginal discs or buds are actually
rudiments of future organs of the adult, such as mouth parts, wings, antennae, walking legs, and
internal organs [Link] discs develop directly from the eggs and remain nonfunctional
throughout the larval stages. During the pupal stage they grow in size and differentiate to form
adult structures which remain collapsed and folded. When the reorganization is completed
the pupa moults to set free the adult or [Link] the adult or imago blood is pumped into
these collapsed structure it causes them to unfold and inflate. Furthermore chitin is deposited on
then to harden them. The mode of development of imaginal disc varies from species to species
and also from organ to organ. Imaginal discs are sometimes formed during late embryonic
development and their cells are separate from the prospective larval cells, as for example in
Drosophila and other Diptera. In some insects the imaginal discs are derived from larval cells
during the later phase of larval growth.
Factors controlling metamorphosis in insects:For a larval moult to be successful all parts of
the body must take part in the process and carry it out at the same [Link] indicates that a
common factor acts on all parts of the body. The existence of a common factor or cause is even
more apparent in metamorphosis, in which the involvement of both external and internal organs
may be more radical and [Link] common factor may be external or internal.
External factors:In some cases an external'factor may be responsible for initiating moulting, as
for instance in the blood sucking Rhodnius the intake of food (blood meal) is such a factor.
Another example in which external factor is essential to initiate a moult is the case of the pupa
moth Platysamia cecropia. After pupation the insect falls into a quiescent state with a reduced
rate of metabolism - diapause - which continues throughout winter. It is essential that during this
time the pupa be exposed to cold,otherwise the diapause is prolonged [Link] diapause
may be broken precociously if the pupa is exposed to cold (3' to 5' C) for at least two [Link]
temporary cooling activates the vital processes in the pupa and on return to a warmer
environment development is completed, the pupa moults and the imago [Link] other
insects factors such as humidity, population density etc., appear to initiate metamorphosis.
However in the majority of insects no external cause of any moult has been detected and moults
follow one another at intervals which appear to be determined entirely by the internal processes
in the animal.
Brain neurosectory cells and their hormones:Kopec was the first to suggest the role of
hormones in controlling metamorphosis. On the basis of his experiments on the larval gypsy
moth (Portheria dispar) he observed that at a particular period the brain releases a substance
into the blood which is essential for pupation and hence for metamorphosis. His findings on the
role of brain hormones in metamorphosis of insects was supported by subsequent workers who
found similar mechanisms in different insect groups. Large secretory nerve cells in the brain of
the insects called neurosecretory nerves were also identified as being responsible for affecting
[Link] secretion of these cells is called activation hormone (AH) or brain hormone (BH).
The activation hormone is comprised of proteins or Lippoproteins. The AH or BH after synthesis
pass along the axons of these cells and end blindlv intn a nair nf storage and release organs
called the corpora cardiaca (CC), located in the posterior brain. The CC releases the active
hormone material called prothoracotropic hormone (PTTH) which is a small polypeptide. This
 prothoracotropic hormone acts upon the prothoracic gland, causing it to secrete the moulting
hormone called 'ecdysone'.Corpus allatum (Singular) and Juvenile hormone.-The corpora allata
are rounded glands attached to the posterior side of each corpus cardiacum forming a compact
body just behind the brain. In some (Hemiptera, higher Diptera) they are fused to form a single
median structure. Experiments by Wiggleworth have shown that the CA secretes a hormone
which determines the character of each larval instar by limiting the degree of diffe entiation
towards the development of the adult. He named this hormone as ju f enile hormone (JH). This
is also known as neotenin (youth substance), (Wigglesworth 1954) and gonadotrophic hormone
(Englemann 1957). The juvenile hormone is similar in structure to terpenes. A large number of
compounds with juvenile hormone activity have been isolated and many have been
synthesized. Some synthetic ones appear more potent than the naturally occurring ones. The
juvenile hormones ensures the occurrence of larval moults and inhibits [Link]
presence or absence determines whether the larva will moult into a larval/pupal stage or into
adult stage. The major natural hormone isolated from the adult male Hyalophora cecropia moth
has the following [Link] has been observed that if an extra corpora allata from a second
stage larva is transplanted to the fourth instar last larval instar then the moult does not produce
a pupa or adult, instead an oversized larva develops. Removal of corpora allata early in larval
life results in premature metamorphosis. Prothoracic gland (PTC) or moulting gland and
ecdysone or moulting [Link] third endocrine gland-prothoracic gland- is an irregular
branching mass of glandular cells located in the prothorax (segment of the thorax that bears the
anterior most of its three pairs of legs), in close association with the tracheal lubes and secretes
the hormone ecdysone which has been isolated and crystallized in pure form from Calliphora
and from pupa of a silkworm. It is a unique water soluble steroid and is related closely to
[Link] studies however show that all a ecdy~oneis converted into P ecdysone
(ecdysterone) after liberation into the haemolymph and that ecdysterone is the active moulting
hormone. Ecdysterone exerts its effect directly upon those cells concerned in growth and
moulting; it activates them and stimulates them to renew protein synthesis. It is believed that it
acts directly upon those loci in the chromosomes which are concerned with growth. It induces
characteristic puffing in the giant polytene chromosomes of Diptera. These puffs are considered
to be the sites for the formation of the messenger RNA needed for protein synthesis.
Metamorphosis is the dramatic biological process by which an insect physically develops,
involving a noticeable change in its body structure. This complex process is precisely controlled
by a cascade of hormones secreted by several endocrine [Link] type of metamorphosis is
determined by the relative concentrations of two key hormones: ecdysone and juvenile hormone
(JH). The endocrine glands and their [Link] endocrine glands and neurosecretory
cells orchestrate insect metamorphosis:

 Neurosecretory cells (NSC) of the brain: These specialized neurons are the master controllers
of metamorphosis. They respond to environmental cues (like light, temperature, and food
availability) and internal signals (like body size) by producing a neurohormone
called prothoracicotropic hormone (PTTH).

 Corpora cardiaca (CC): These are paired, neurohaemal organs attached to the brain. They act
as storage and release centers for PTTH produced by the neurosecretory cells.

 Prothoracic glands (PG): Located in the prothorax, these glands are activated by PTTH. In
response, they secrete the steroid hormone ecdysone, which is also known as the molting
hormone. Ecdysone is later converted into its active form, 20-hydroxyecdysone (20E), in
peripheral tissues.

 Corpora allata (CA): These paired glandular bodies are located behind the brain. They are
responsible for producing and secreting juvenile hormone (JH).
 The hormonal cascade controlling molting and metamorphosis
The developmental pathway of an insect is determined by the interaction between ecdysone
(which promotes molting) and juvenile hormone (which prevents the transition to adult stages).

1. The larval stage (high JH)

 Throughout the juvenile larval instars, both ecdysone and juvenile hormone are present in the
hemolymph (insect blood).

 Mechanism: Neurosecretory cells in the brain release PTTH, which stimulates the prothoracic
glands to secrete ecdysone. At the same time, the corpora allata are highly active, secreting a
high concentration of juvenile hormone.

 Outcome: The presence of a high titer of juvenile hormone during a molt prevents the
expression of adult characteristics. The insect sheds its old exoskeleton and molts into a larger
version of its current larval stage.

2. The pupal stage (low JH)

 When the larva reaches a genetically predetermined critical size, it begins preparing for the
metamorphic molt.

 Mechanism: The neurosecretory cells signal a reduction or cessation of juvenile hormone

secretion from the corpora allata. The corpora allata atrophy and become inactive. As a result,
the juvenile hormone levels in the hemolymph drop significantly. With the high ecdysone levels
still present, the absence of high JH concentration is the critical signal for metamorphosis.

 Outcome: The larva molts into a pupa, which is a transitional, non-feeding stage in
holometabolous insects (those with complete metamorphosis).

3. The adult stage (no JH)

 The adult stage is the final transformation that occurs inside the pupal cuticle.

 Mechanism: During the final molt from pupa to adult, there is no juvenile hormone present in the
insect's system. The ecdysone, acting alone, triggers the final differentiation process.

Outcome: The adult insect (imago) emerges with fully developed adult characteristics, such as
functional wings and reproductive organs It is well established that Metamorphosis or the post
embryonic growth of insects are controlled and regulated by neurosecretions and hormones .
Certain nerve cells are modified to secrete [Link] nerve cells are called
neurosecretory cells and their secretions are called neurosecretions . The neurosecretions are
temporally stored in special structures called corpus cardiacum .
Neurosecretions in Insects :-The various hormones secretedby neurosecretory cells of the
brain are as follows :
1. Brain hormone ( BH ):Brain hormone is secreted by the neurosecretory cells of the brain .
Chemically it is a [Link] hormone serves to activate the corpora cardiaca,a component of
the retrocerebral complex of the stomatogastric nervous system .
2. Prothoracicotropic hormone (PTTH ) :This hormone is secreted by the corpora cardiaca ,
which in turn stimulates the prothoracic glands .
3. Prothoracic gland hormone ( PGH ):This hormone is secreted by the paired , bilateral sheet
of cells in thorax,constituting the prothoracic [Link] it is [Link] hormone is
known to trigger moulting as it acts on the tissues to promote all of the changes characterizing a
Moult.
.
[Link] hormone ( JH ):This hormone is secreted by another component of the
retrocerebral complex , the corpora [Link] it is an unsaponifiable , non-sterolic lipid .
This hormone regulates morphogenesis and so promotes metamorphosis , that is , development
of the larvae into adult through pupal stage .
Conclusion :-During metamorphosis , the adult character develop in the form of buds called
imaginal discs . They are the future [Link] imaginal discs remain folded in the pupa . They
increase in size and differentiate into adult [Link] , hormonal control of metamorphosis
can be summarized as follows : a) The brain secretes a hormone called brain hormone . It
stimulates an endocrine gland called prothoracic gland .b) The prothoracic gland secretes a
hormone called ecdyson . The ecdyson induces moulting ,growth & differentiation . This
hormone converts the larvae into the adult .c) Corpora allata an endocrine gland attached to the
brain secretes another hormone called Juvenile hormone . It keeps the larva in
the larval stage . As long as it remains in thebody , the larva cannot become an adult . When
this hormone decreases , the larva becomes an adult .
MOULTING
Metamorphosis in biology means the process of transformation from an immature form to an
adult form in two or more distinct stages. Good examples are insects and [Link] for
most insects begins as a larva or nymph then progresses to the pupa stage and ends as an
adult. Animals that metamorphosize include tadpoles into frogs, caterpillars into butterflies, bee
larva into bees, and [Link] life cycle is generally complex involving several stages of the
larval and pupal development. Adults are generally quite different from the larval forms. When
the larvae undergo considerable change to become adults it is called [Link]
physical transformation of an insect from one stage of its life cycle to another is called
[Link] process of producing the new cuticle and subsequent shedding (ecdysis) of
the old cuticle is called [Link] accompanied by a change in a body form is known as
 [Link] degree of metamorphosis depends on the degree of divergence between
immature and adults.
PROCESS OF MOLTING:
Two main process in moulting: It mainly involve two distinct process which may be widely
separated in time so that it is convenient to distinguish between them. The two main process
are 1. Apolysis - separation of old cuticle 2. Ecdysis - shedding of remnants of the old cuticle
STEPS INVOLVED IN MOULTING PROCESS: 1. Apolysis 2. Secretion of inactive moulting
fluid by epidermis 3. Production of cuticulin layer of new exoskeleton 4. Activation of moulting
fluid 5. Digestion and absorption of old endocuticle 6. Epidermis secretes inner epicuticle and
new procuticle 7. Ecdysis 8. Expansion of new integument 9. Tanning
Changes in epidermis: The onset of moulting is indicated first by changes in the
epidermal cells which divide mitotically and become columnar in form; they are flattened
the over all area of the epidermis, hence the cuticle size is increased • Separation of old
cuticle from the epidermis - As a result of changes in cell shape, a tension is generated
at the surface of the epidermal cell which result in their separating from the cuticle •
Digestion of old endocuticle - As the cuticle separate from the epidermis moulting fluid is
secreted into the space. As a result of activity of moulting fluid line of weakness appears
in the ecdysial line. The moulting fluid contain the enzyme proteinase and chitinase
ECDYSIS • Usually ecdysis follows as soon as digestion is complete • As a process of
separating the old cuticle which consist of epicuticle and exocuticle from the new cuticle • In
sometime the old cuticle may retain for a time and the insect referred to as pharate instar
DURING ECDYSIS • The pore canal within the procuticle allow the movement of lipids and
proteins towards the new epicuticle where the wax layer and cement layer form • Forms shortly
before ecdysis wax is separated on to the surface of the new cuticle and the layer adjacent to
the cuticulin forms oriented monolayer • Soon after ecdysis the layer formed is the cement layer.
TANNING • The wrinkled new integument is expanded by stretching • Differentiation of
procuticle into exocuticle and epicuticle take place(after ecdysis) by sclerotization of exocuticle
alone • After hardening muscle relax ,and air is exhaled ,blood pressure reduce and the space is
now available for further protoplasmic synthesis • Until the process of tanning is complete it is
called as in teneral condition.
Environmental Factors:
Temperature: Temperature plays a significant role in the timing of insect
development, including the rate of moult. Insects have specific temperature
thresholds for development.
 Photoperiod (Light): Changes in light levels can also influence hormonal activity
and trigger molting.
 Nutrition:Adequate food sources are essential for insect growth & development,as
insect needs enough energy to initiate & complete the energetic process of molting.
 Humidity: Humidity is an important factor affecting insect survival and development,
and can influence the success of the molting process.
 Population Density: High population densities can lead to increased competition
for resources, which can affect nutrition and, indirectly, molting and development.
The Process of Ecdysis:
1. Hormonal Trigger: A pulse of 20-hydroxyecdysone initiates moulting.
2. Epidermal Changes: Epidermal cells divide, enlarge, and separate from the old cuticle
(apolysis).
3. Moulting Fluid: Moulting fluid is secreted to digest the old endocuticle.
4. New Cuticle Formation: A new, soft cuticle forms beneath the old one.
5. Ecdysis: The insect emerges from the old cuticle.
6. Cuticle Hardening and Darkening: The new cuticle expands, hardens, and darkens to protect
the insect.
Molting, known technically as ecdysis, is literally a period of growth for insects. Insects grow in
increments. Each stage of growth ends with molting the process of shedding and replacing the
rigid exoskeleton. People often think molting is the simple act of an insect breaking out of its
skin and leaving it behind. In truth, the process is complex and involves several [Link] egg
hatches, the immature insect feeds and grows. Its exoskeleton is like a shell. Eventually, the
larva or nymph must shed its unyielding overcoat to continue its development. The exoskeleton
which serves as its external backbone is used for protection and support. Without an
exoskeleton, the insect could not survive. An old exoskeleton is shed when a new one is ready
underneath, a process that can take days or [Link] undergo the process of molting, an
insect must begin to take in air or water by either swallowing it in naturally or raising its internal
blood pressure. This instigates the process of molting that begins. The result is a soft,
expandable exoskeleton suitable for further growth. This process is repeated several times
during the life span of an insect depending on the species. The new exoskeleton will eventually
harden and retain the original colouring of the insect as it matures and is exposed to the
elements and every day wear-and-tear. In molting, the epidermis separates from the outermost
cuticle. Then, the epidermis forms a protective layer around itself and secretes chemicals that
break down the insides of the old cuticle. That protective layer becomes part of the new cuticle.
When the epidermis has formed the new cuticle, muscular contractions and air intake cause the
insect’s body to swell, thus splitting open the remains of the old cuticle. Finally, the new cuticle
hardens. The insect must continue to swell and expand the new cuticle, so it is large enough to
allow room for more growth. The new overcoat is soft and much paler than the former one, but
over a few hours, it becomes darker and begins to harden. Within a few days, the insect
appears to be a slightly larger copy of its former self.
Hormonal control of Insect Metamorphosis: The hormonal control of insect metamorphosis
was shown by Wigglesworth (1934), who studied Rhodnius prolixus, a blood-sucking bug that
has five instars. When the 1st instar larva of Rhodnius was decapitated and fused with the 5th
instar larva, the minute first instar developed the cuticle, body structure, and genitalia of the
adult. Wigglesworth also showed that corpora allata located near the insect brain, produce a
hormone that slow down metamorphosis by producing juvenile hormone. This hormone inhibits
the genes that promote development of adult characteristics (e.g. wings, reproductive organs,
and external genitalia), causing the insect to remain nymph or [Link] molting process is
initiated in the brain, where neurosecretory cells release prothoracicotropic hormone (PTTH) in
response to neural, hormonal, or environmental factors. PTTH stimulates the production of
ecdysone by the prothoracic [Link] second major hormone in insect development is
juvenile hormone (JH). JH is secreted by corpora allata. The secretory cells of corpora allata are
active during larval molts but are inactive during the metamorphic molt. This hormone is
responsible for preventing metamorphosis.
When an immature insect has grown sufficiently to require a larger exoskeleton, sensory input
from the body activates certain neurosecretory cells in the brain. These neurons respond by
secreting brain hormone which stimulates corpora cardiaca to release the prothoracicotropic
hormone (PTTH) into circulatory system, which then stimulates the prothoracic glands to
secrete the molting hormone, [Link] hormone acts as the developmental switch
between immature and adult forms, a sesquiterpenoid produced by the corpora allata gland. As
long as the juvenile hormone is present in sufficient concentration, promotes the
growth of larva, moults result in another larval instar but inhibits the differentiation of adult
structures. In the last larval instar, an axonal nerve from the brain inhibits the corpus allatum
from releasing juvenile hormone. The lowered JH level allows metamorphosis to continue.
20-hydroxyecdysone (20E) promotes the successive molts, including the metamorphic one,
whereas JH is a terpenoids (lipid), which inhibits metamorphosis. 20-hydroxyecdysone initiates
and coordinates each molt and regulates the changes in gene expression, induces ecdysis,
which mean transfer one stage to another (metamorphosis). Each molt is initiated by one or
more pulses of 20-hydroxyecdysone. For a larval molt, the first pulse produces a small rise in
the hydroxy ecdysone concentration in the larval hemolymph (blood) and elicits achange in
cellular commitment. A second, large pulse of Hydroxy ecdysone initiates the differentiation
events associated with [Link] hydroxyecdysone produced by these pulses commits and
stimulates the epidermal cells to synthesize enzymes that digest and recycle the components of
the cuticle. Juvenile hormone prevents the ecdysone-induced changes in gene expression (from
immature to mature) that means prevents the development of adult characteristics. The
presence of juvenile hormone during a molt ensures that the result of that molt produces
another instar, not a pupa or an adult.
Tissue reactivity:
The mechanism of metamorphosis in anurans indicates that a single agent, namely thyroxine,
evot?.: rr.;rltiple responses in different tissues. In other words, the variousorgans of the body
respond differently to a single hormone agent. In addition, the response of the tissues, though
diversified, are specific and can be destructive or constructive depending on the target tissue.
Thus the same stimulus will cause certain tissues to degenerate and others to grow and
[Link] diverse reaction of tissues is believed to be due to two reasons
(1) Competence of larval tissue or multiple response. The responsiveness of different larval
tissues to thyroxine is markedly different. The tissues of the tail becomes necrotic and undergo
degeneration due to the action of the thyroid hormone,while the tissues of the limb increase and
undergo [Link] thyroxine causes rapid aging and destruction of R.B.C's of
the larva undergoing [Link] addition it stimulates the development of cells that
synthesize the haemoglobin of adult frog. The hormone T3 reduces biosynthesis of nucleic
acids in the tail but increases their synthesis in the liver. Furthermore, muscles of the tail
undergo degeneration while those of the trunk remain [Link] examples clearly
indicate that the response and reactivity of larval tissue to the same hormone or agent differs
greatly. In addition it can also be demonstrated experimentally that the reactivity and response
of the target tissues are intrinsic,specific and [Link] can be demonstrated by
transplanting tail tips of the tadpole to the trunk of another tadpole undergoing metamorphosis,
or by placing the eye of the tadpole in the tail of a metamorphosing larva (Schwind. 1933,
Geigy. 1941).The extra tail transplanted into the tadpole host trunk is not protected from
degeneration and undergoes necrosis along with the host tail and gets absorbed. The eye
retains its integrity despite the fact that it lies surrounded within the degenerating tail. The
degeneration of tissues during metamorphosis thus represents genetically determined cell
death. In humans such programmed degeneration occurs in the tissues between our fingers and
toes. The degeneration of the human tail during the week four of development resembles the
regression of tadpole [Link] observations force us to question as to how does the thyroid
hormones bring about diverse metamorphic changes? Do the hormones act directly on all the
target tissues or are their actions mediated by some other hormone/honnones? Finally, how is
the changing thyroxine tiues controlled? Such questions have been answered by experiments
which have clearly shown that the hormone acts directly on the target tissue. Weber (1967)
demonstrated this fact when he cultured excised tadpole tails in the presence of [Link] tails
treated with thyroxine showed regression similar to that occuning in metamorphosis, the
controls (not treated wilh thyroxine) exhibited no regression and remained healthy. The
regression of the tail is believed to occur in four stages. First, protein synthesis decreases in the
treated muscle cells of the tail. Then, there is an increase in the lysosomal enzymes.
Concentrations of Cathepsin D (a protease), RNase, DNase collagenase, phosphotase, and
glycosidases all rise in the epidermis, notochord and nerve cord cells. Cell death is probably
caused by the release of these enzymes into the cytoplasm. The epidermis helps to digest the
muscle tissue, probably by releasing these digestive enzymes. After this death, macrophages
collect in the tail region,digesting the debris with their own proteolytic [Link] result is that
the tail becomes a large sac of proteoltic [Link] the epidermis is removed from the tail tips,
the tips will not regress when cultured in thyroxine.
2. Threshold value of different larval tissues for the thyroid hormone concentration
One of the major problems of metamorphosis is the coordinalion of developmental
events. The tail should not degenerate till some other means of locomotion, namely the limbs
develop, and the gills should not regress till the animal can use its newly developed lung
[Link] (1961) demonstrated the possibility of this coordination which shows that
different larval tissues have different threshold value for thyroxine. In other words the different
larval tissues are sensitive to different concentrations of thyroid [Link] model is called
the threshold [Link] has been observed that structures in the tadpole which change early
during metamorphosis are more sensitive to and have low threshold value to thyroxine, than
those which undergo transformation at a later stage. As metamorphosis commences the thyroid
hormone level gradually builds up and different events occur at different concentrations of the
hormone. Experimental studies have revealed that tadpole pans which have a low threshold
respond earlier during metamorphoses than those parts having a high threshold response. In
other words, the threshold value reflects the order in which metamorphic changes occur in
normal [Link] urodeles the bulging of eyes react to the weakest dose of thyroid
hormone (minimum threshold) and so is the first event in metamorphosis. This is followed by
reduction of fin fold and the shortening and disappearance of external [Link] which occurs
the closure of gill clefts and transformation of [Link] administration of high amounts of
thyroxine at early stages causes precocious but distorted sequence of metamorphic changes
resulting in death. Tissues. responsive to low concentrations will not respond, while those
responsive to high amount do so precociously. For example; in the frog tadpoles as hormone
concentration increases tissue responses become progressively more rapid till maximum rates
of change are attained. When the dosage is heavy all metamorphic events start together or
crowd together and the normal sequence of events is disturbed. The destructive processes
being capable of proceeding faster than the constructive processes result in the forelimb
erupting before becoming differentiated; the tail becomes reduced before the legs are
sufficiently developed to take over locomotion-resulting in an abnormal animal which dies. There
is a progressively increasing threshold of the hormones at different stages of development of a
particular organ and during the entire metamorphic process.
Induction in metamorphosis
Some of the morphegenetic changes during metamorphosis are found fo be quite independent
of hormone action. For example, usually during metamorphosis, the skin of the tail undergoes &
generation under hormonal effect. It is observed that when the skin alone is removed from tail
and transplanted onto the tail of another metamorphosing larva, it does not regress despite the
presence of the hormone. But if the skin is grafted to the trunk along with its underlying muscle
in a developing tadpole,then it regresses. Thus the hormone affects only the muscle directly,
which induces regression or progression of the skin depending upon the induction it receives
from its underlying muscle of tail or trunk respectively. Similarly tympanum is another example
of the induction process and is independent of direct hormone, action as its formation is induced
by the tympanic cartilage.
PATTERNS OF EGGS:Eggs are classified mainly on the basis of
1. On the Basis of the Amount of Yolk
i. Alecithal Egg:When the egg contains no yolk, it is called alecithal [Link]. The eggs of
eutherian mammals
ii. Microlecithal Egg:When the egg contains small or negligible amount of yolk, it is said to be
microlecithal .Romer (1962) and Balinsky (1970) named these eggs as oligolecithal eggs. Eg.
Amphioxus

iii. Macrolecithal or Megalecithal Egg:When the egg contains large amount of yolk, it is said
to be macrolecithal or megalecithal [Link] is also called polylecithal egg. Eg. Bony fishes,
amphibians, reptiles, birds, [Link] amphibians, Dipnoi and Petromyzon, the amount of yolk
present is not very high. Hence these eggs are also named as mesolecithal eggs

2. On the Basis of the Distribution of Yolk

i. Homolecithal or Isolecithal:In isolecithal egg, the yolk materials are uniformly distributed
throughout the eggs. [Link] and Amphioxus
ii. Telolecithal :Here the yolk is highly concentrated towards the vegetal [Link] amount of
yolk gradually decreases from the vegetal pole towards the animal pole. Eg. Fishes,amphibians,
reptiles and birds
iii. Centrolecithal:In centrolecithal egg, the yolk takes up a central position and is
surrounded by a thin layer of cytoplasm. Eg. Insects
3. On the Basis of the Presence or Absence of Shell
i. Cleidoic Egg:The eggs with hard shell are called cleidoic eggs. Eg. Reptiles, Birds, etc

ii. Non-cleidoic Egg:The non-cleidoic eggs are not protected by [Link] type of egg is in
animals in whose case the development is internal. Eg. Mammals
Polarity
The unequal distribution of egg components creates the animal pole and vegetal pole in the
egg, it is called polarity.

Gradient
A gradient is defined as a proportional rise and fall of certain factors in the embryo, the
interaction of which brings about normal development.
1. The process leads to formation of germ cells or gametes.
2. The normal body cells known as somatic cells are diploid (2n) where as the germ cells are
haploid (n).
3. During fertilization one halpoid sperm unites with one haploid ovum to form a normal diploid
somatic cell thus keeping the chromosome number constant generation after generation.
4. During first maturation division, the reshuffling of paternal and maternal genes take place
resulting in variation.
1. Monospermy Generally, only one spermatozoon enters the egg and fuses with [Link] a
fertilization is said to be [Link] is common in majority of animals like
coelenterates, annelids, echinoderms, bony fishes, frogs, etc
2. Polyspermy In some animals, normally many sperms enter the [Link] a fertilization is
said to be [Link] many sperms enter the egg, only one sperm nucleus fuses
with the egg nucleus. Other sperm nuclei [Link] naturally occurs in animals
with heavily yolked [Link]. Arachnids, some insects, elasmobranchs, urodeles, reptiles,
birds, etc. This is known as physiological polyspermy
3. Acrosome Reaction When the spermatozoon comes in contact with the egg, tremendous
changes occur in the acrosome of [Link] these changes constitute the acrosome reaction.
The acrosome reaction has been studied extensively in a variety of animals. Colwin (1967)
demonstrated the acrosome-reaction in the enteropneust Saccoglossus .When the
spermatozoon’s tip makes contact with the egg envelope, the sperm-plasma membrane and the
acrosomal membrane rupture at the point of contact .The acrosomal membrane then joins with
the plasma membrane around the margin of the [Link] acrosomal granule is released
on the egg-envelope .The acrosomal granule contains the sperm lysin which dissolves the egg
envelope. Thus the egg surface is exposed at this [Link], the centre of acrosomal
membrane, nearer to the nucleus, grows towards the egg surface as a thin tube called
acrosomal [Link] the acrosomal tube grows further, its tip comes in contact with the plasma
membrane of the [Link] some cases, more than one acrosomal tubes develop. Eg. Hydroides
(Polychaete)
Morphogenetic process
[Link] by differential growth: After their initiation, the various organs and regions
of an organism may increase in size at different rates. Such processes of differential growth will
change the overall shape of the body in which they occur. Processes of this kind take place very
commonly in animals, particularly in the later stages of development. They are of major
importance in the morphogenesis of plants, where the overall shape of the plant, the shape of
individual leaves, and so on, depends primarily on the rates of growth of such component
elements as the stems, the lateral shoots, and the vein and intervein material in leaves. In both
animals and plants, such growth processes are greatly influenced by a variety of hormones. It is
probable that factors internal to individual cells also always play a role. Although differential
growth may produce striking alterations in the general shape of organisms, these effects should
probably be considered as somewhat superficial, since they only modify a basic pattern laid
down by other processes. In a plant, for instance, the fundamental pattern is determined by the
arrangement of the lateral buds aroundthe central growing stem; whether these buds then grow
fast or slowly relative to the stem is a secondary matter, however striking its results may be.
II. Morphogenetic fields: Many fundamental processes of pattern formation (e.g., the
arrangement of lateral buds in growing plants) occur within areas or three-dimensional masses
of tissue that show no obvious indications of where the various elements in the pattern will arise
until they actually appear. Such masses of tissue, in which a pattern appears, have been
spoken of as “fields.” This word was originally used in the early years of the 20th century by
German authors who suggested an analogy between biological morphogenetic fields and such
physical entities as magnetic or electromagnetic fields. The biological field is a description, but
not an explanation, of the way in which the developing system behaves. The system develops
as though each cell or subunit within it possessed “positional information” that specifies its
location within the field and a set of instructions that lays down the developmental behaviour
appropriate to each position. There have been several attempts to account for the nature of the
positional information and of the corresponding instructions. The oldest and best known of these
is the gradient hypothesis. In many fields there is some region that is in some way “dominant,”
so that the field appears as though organized around it. It is suggested that this region has a
high concentration of some substance or activity, which falls off in a graded way throughout the
rest of the field. The main deficiency of the hypothesis is that no one has yet succeeded in
identifying satisfactorily the variables distributed in the gradients. Attempts to suppose that they
are gradients of metabolic activity have, on investigation, always run into difficulties that can
only be solved by defining metabolic activity in terms that reduce the hypothesis to a circular
one in which metabolic activity is defined as that which is distributed in the gradient. Recently, a
new suggestion has been advanced concerning position information. Most processes within
cells normally involve negative feedback control systems. These systems have a tendency to
oscillate, or fluctuate regularly. In fact, any aspect of cell metabolism may be basically
oscillatory in character; the cycle of cell growth and division may be only one example of a much
more widespread phenomenon. The substances involved in these oscillations are likely to
include diffusible molecules capable of influencing the behaviour of nearby cells. It is easy to
envisage the possibility that there might be localized regions with oscillations of higher
frequency or greater amplitude that act as centres from which trains of waves are radiated in all
directions. It has been suggested that positional information is specified in terms of differences
in phase between two or more such trains of transmitted oscillations. Certain types of field
phenomenon may involve an amplification of stochastic (random) variations. In systems
containing a number of substances, with certain suitable rates of reaction and diffusion, chance
variation on either side of an initial condition of equilibrium may become amplified both in
amplitude and in the area involved. In this way, the processes may give rise to a pattern of
differentiated areas, distributed in arrangements that depend on the boundary conditions.
III. Morphogenesis by the self-assembly of units: Complex structures may arise from the
interaction between units that have characteristics such that they can fit together in a certain
way. This is particularly appropriate for morphogenesis at the simple level of molecules or cells.
Units such as the atoms of carbon, hydrogen, oxygen, nitrogen, and so on, can assemble
themselves into orderly molecular structures, and larger molecules, such as those of
tropocollagen, or protein subunits in general, can assemble themselves into complexes whose
structure is dependent on localized and directional intermolecular forces. It seems that such
comparatively large entities as the units that come together to form the head structures of
bacteriophages or bacterial flagella are capable of orderly self-assembly, but the chemical
forces that give rise to the interunit bonds are still little understood. Processes that fall into the
same general category as self-assembly may occur within aggregates of cells. The units that
self-assemble are the cells themselves. Interaction and aggregation may be allowed to occur in
assemblages of cells of one or more different kinds. In such cases it is commonly found that the
originally isolated cells tend to adhere to one another, at first more or less at random and
independently of their character, but later they become rearranged into a number of regions
consisting of cells of a single kind. When the cells in the initial collection differ in two different
characteristics, for instance in species and organ of origin, the assortment in some cases brings
together cells from the same organ, in other cases cells from the same species. Mixtures of
chick and mouse cells, for instance, reassort themselves into groups derived from the same
organ, whereas cells from two different species of amphibia sort out into groups from the same
species more or less independently of organ type. This morphogenetic process probably has
only a restricted application to the formation of structures in normal development, in which only
in a few tissues (e.g., the connective system) do cells ever pass through a free stage in which
they are not in intimate contact with other cells, and cells of different origin do not normally
become intermingled so as to call for processes of reassortment. To explain normal
morphogenetic processes of plants and animals one must look to the results that can be
produced by the differential behaviour of cells that remain in constant close contact with one
another. Several authors have shown how striking morphogenetic changes could be produced
within a mass of cells that remain in contact, but that undergo changes in intensity of adhesion
between neighbouring cells, in the area of surface in the proportion to cell volume, and so on.
IV. Differentiation: is simply the process of becoming different. If, in connection with biological
development, morphogenesis is set aside as a component for separate consideration, there are
two distinct types of differentiation. In the first type, a part of a developing system will change in
character as time passes; for instance, a part of the mesoderm, starting as embryonic cells with
little internal features, gradually develops striated myofilaments, and with a lapse of time
develops into a fully formed muscle fibre. In the second type, space rather than time is involved;
for instance, other cells within the same mass of embryonic mesoderm may start to lay down an
external matrix around them and eventually develop into cartilage. In development,
differentiation in time involves the production of the characteristic features of the adult tissues,
and is referred to as histogenesis. Differentiation in space involves an initially similar
(homogeneous) mass of tissue becoming separated into different regions and is referred to as
regionalization. Histogenesis involves the synthesis of a number of new protein species
according to an appropriate timetable. The most easily characterized are those proteins formed
in a relatively late stage of histogenesis, such as myosin and actin in muscle cells. The
synthesis of proteins is under the control of genes, and the problem of histogenesis essentially
reduces to that of the genetic mechanisms that direct protein synthesis. Regionalization is
concerned with the appearance of differences between various parts of what is at first a
homogeneous, or nearly homogeneous, mass. It is a prelude to histogenesis, which then
proceeds in various directions in the different regions so demarcated. The processes by which
the different regions acquire distinct contrasting characteristics must be related to some of the
processes discussed under morphogenesis. Unlike morphogenesis, regionalization need not
involve any change in the overall spatial shape of the tissues undergoing it. Regionalization falls
rather into the type of process for which field theories have been invoked.
Characters of a cleidoic egg:A cleidoic egg is a self-contained reproductive unit with a
protective, sealed shell. This key adaptation allows for reproduction on land, away from water,
and is characteristic of terrestrial animals like birds, reptiles, and some insects.
[Link] shell: A hard, outermost calcareous shell provides protection from physical
damage and desiccation (water loss). The shell can be either hard, as in birds, or leathery and
flexible, as in many reptiles.

[Link] to gases: While the shell prevents water from escaping, it is porous and allows
the exchange of gases like oxygen and carbon dioxide. This is crucial for the embryo's
respiration.

[Link]-contained nutrition: The egg contains a large amount of yolk, which serves as a
complete food source for the developing embryo. This eliminates the need for a larval stage that
must feed independently.

[Link] membranes: Specialized membranes support the embryo's development

within the [Link]: The innermost membrane, which encloses the embryo in a fluid-filled
cavity. It protects the embryo from shock and prevents drying [Link]: The outermost
membrane, which lies just beneath the eggshell and facilitates gas [Link]: A sac-
like membrane that functions as a respiratory organ and holds the embryo's metabolic waste
(especially uric acid, which conserves water).

[Link] adaptation: The cleidoic egg is a critical evolutionary innovation that enabled
vertebrates to invade and thrive in terrestrial environments. It provides a private, protective pond
for the embryo, freeing it from the constraints of an aquatic environment.

[Link] fertilization: Since the egg is sealed before being laid, internal fertilization is a
necessary prerequisite for laying a cleidoic egg.

Structure of a typical cleidoic egg:The chicken egg is a classic example of a cleidoic egg,
with the following key components:
Eggshell: The hard, outermost covering made of calcium carbonate. It is permeable to gases
but prevents [Link] membrane: A double membrane just inside the shell. At the
blunt end of the egg, it separates to form the air [Link] (egg white): A protein-rich
layer that surrounds the yolk. It provides water and further cushioning for the [Link]: The
primary nutrient source for the [Link]: Rope-like structures that anchor the yolk to the
shell membrane, keeping it centrally [Link] disc (blastoderm): A small, whitish
spot on the surface of the yolk. It contains the egg nucleus and is where embryonic
development begins after fertilization.
 TERATOGENESIS
Prenatal toxicity characterized by structural or functional defects in the developing embryo or
fetus is known as Teratogenesis. It too includes intrauterine growth retardation, death of the
embryo or fetus, and transplacental carcinogenesis (in which chemical exposure of the mother
initiates cancer development in the embryo or fetus, resulting in cancer in the offspring after
birth). Intrauterine human development has three stages: implantation, post-implantation, and
fetal development. The first two stages of implantation and post-implantation are the embryonic
stages and last through the first eight weeks after conception. The fetal development stage
begins in the ninth week and continues to birth. The developmental stage depending on
chemical contact with the mother can result in different degrees of toxicity in the embryo or
fetus. In the preimplantation phase, a toxic chemical can kill some of the cells in the blastocyst,
resulting in the death of the embryo. Throughout the postimplantation period, chemical-induced
cell death leads to one of two outcomes. If death is controlled by those cells undergoing active
cell division at the moment, the analogous organs are affected, resulting in malformation. If the
cell death is widespread with no significant replication by the remaining cells to sustain life, the
embryo dies. During the third, fetal, period, chemical damage can retard growth or, if severe
sufficient, kill the fetus. A congenital malformation is a gross structural present at birth. Its
incidence is about 2.5% in all infants born. But, only half of these deformities are noticeable at
the time of delivery, most of the remainder coming to light during the first postnatal year. The
term congenital anomaly is reserved for a minor congenital disorder such as a deformed finger
or ear lobe. Anomalies are found in a further 2.5 percent of live-born infants. Individuals of a
species (including humans and other animals) exhibiting minor deviation variation from each
other are considered normal. Individuals that show gross deviation from each other are
considered normal due to congenital malformations and are known as monsters or terata. The
abnormal development or formation of terata is called teratogenesis and the science which is
concerned with the investigation of terata and teratogenesis is called teratology. Teratogenesis
is the formation of an abnormal organism. A teratogen is any manager that physically or
chemically alters developmental processes and produces inherited deformities. The nature of
the teratogen and the developmental stage during which the variation occurs is critical to the
type and severity of abnormality it will produce. Biological factors such as the organism’s
gestation process, developmental pathways, and life-cycle characteristics also control the
exposure and effect of a teratogen. Mechanical disruptors, environmental factors, and chemical
contaminants are the primary categories of teratogens disturbing wildlife [Link]
introduction of an embryo, fetus, or larva to a teratogen may result in death, structural
malformation, functional disorder, or growth retardation. The most commonly described
teratogenic effects taking place in ecosystems are external malformations. Wild organisms have
always been subject to teratogenic insult; however, current anthropogenic activities have
increased the prevalence of deformities. Herein, the current state of teratogenesis knowledge
concerning amphibians, reptiles, birds, fishes, mammals, and invertebrates is discussed with
emphasis on structural malformations resulting from exposure to chemical contaminants.

TYPES OF TERATOGENESIS:Teratogenesis is Genetic teratogenesis and environmental

teratogenesis. Genetic teratogenesis has been studied both in human beings and animals.
Genetic teratogenesis in human beings: Abnormal genes may be inherited from one or both
parents and they may be dominant or recessive. In the majority of the afflicted individuals,
however, a Mendelian pattern of inheritance cannot be observed, the abnormalities merely
occur more frequently among relatives than in the general population. In those occurring among
relatives, there may be a complex interplay between several genes. Deformities due to
abnormal dominant genes are rare. In most of these, the skeleton is affected and the
deformities include achondroplasia (insufficient growth of long bones), and arachnodactyly
(abnormally long hand and foot bones). A teratogen is a substance that may lead to birth
defects in an embryo or fetus. During pregnancy, contact with certain chemicals, infections, and
drugs may increase the risk that a person will miscarry or that the embryo or fetus could have a
developmental abnormality. Alcohol and smoking are two common teratogens. Contact with
either of them can lead to developmental anomalies, miscarriage, stillbirth, preterm labor, and a
variety of other pregnancy complications. The contact of teratogens with pregnancy or a fetus
depends on numerous factors. The timing and length of exposure, the stage of pregnancy when
the exposure happened, whether a parent’s genes make them more at risk, and the type of
agent they were exposed to all contribute to the risk. Teratogens commonly fall under the
following categories: 1. Drugs 2. Infection 3. Physical Agents 4. Environmental Toxins 5.
Maternal Health Conditions.

Genetic teratogenesis in animals: Genetic teratogenesis can be considered under the

following two headings: 1. Gene-phene relationship. Several different genes can cause the
same terata, though not necessarily by the same route. For example, there are more than
twenty genes that affected eye color in Drosophila melanogaster. The mutants causing the
same defect may be either recessive or dominant. For example, in fowl, the trait of
rumplessness (absence of tail) is controlled by either a recessive or dominant gene. In some
cases, the same mutation may behave as recessive or dominant depending on the genetic
background. Thus, in mice, the fused gene (for fusion or absence of ribs and or absence of tail)
is dominant in Mus musculus but recessive in Mus musculus bactrianus. The penetrance (i.e.,
the proportion of affected individuals in a population) and expressivity (i.e., degree of effect) of
mutant genes is dependent on both genetic and environmental factors. For example, fowl
carrying rumples gene can be selectively bred to produce a ‘normal’ tail phenotype. Similarly, in
Drosophila carrying the Bar eye gene, the size of the eye and the number of facets in the eye
decreases by about 100 facets with an increase in temperature (15° to 31°C) during the
development. 2. Autophene, allophone, and pleiotropy. Not all genetic terata are the result of
the intrinsic action of genes in the affected tissue. The following two examples will make the
point clear: (1) Creeper mutation (cp/cp) in fowl affects the limbs (called phocomelia or
abnormally short limbs) and the eyes (called microphthalmia or small eye), the embryo does not
survive till hatching. Transplants of the cp/cp limb rudiments in the normal hosts produce
phocomelia limbs. However, transplants of cp/cp eye rudiments in the normal hosts produce
normal eyes. Therefore, the cp gene intrinsically affects limb development (it is called
autophene) but only indirectly affects eye development (it is called allophone). (2) The multiple
effects of one gene (pleiotropy) are also now better understood. Any given gene mutation
essentially affected the production or structure of one transcribed RNA molecule. The
translation product of this RNA (i.e., mRNA), the defective protein, may ultimately result in
various defects due to the correlation of various biochemical reactions in the body (i.e., by a
pedigree of causes), e.g., death in rats due to grey lethal mutant gene and sickle cell anemia in
human beings.

Environmental teratogenesis: Almost any environmental factor can be teratogenic. The

environmental factor may be either biological (e. g., viruses) or non-biological such as physical
factors (e.g., temperature, irradiation, mechanical disturbance) and chemical factors (e.g.,
drugs, environmental chemicals, and dietary imbalances or malnutrition). 1. Teratogenesis due
to infection: The most dangerous known teratogenic organism is the virus of German measles
(rubella). Contraction of the disease by the mother during the first month of pregnancy appears
to carry about a 50 percent risk of producing a congenital abnormality, during the second month
the risk is about 25 percent, and during the third month about 7 percent. The virus tends to
affect the developing eyes, ears, and palate. The triads of congenital cataracts (blindness),
heart disease, and deafness have become known as the rubella syndrome. Rubella also knows
to cause terata such as mental deficiency due to a very small brain, cleft palate, and hare-lip.
Cytomegalovirus (CMV) infection is found all over the world. Like the rubella virus, it passes
through the placenta to infect the fetus. The CMV is show signs of disease. It may cause
deafness, mental retardation, epilepsy, liver disease or cerebral palsy (paralysis due to damage
to the brain), or a variable combination of these. Other teratogenic organisms are toxoplasmosis
& [Link] is known to cause stillbirth or other congenital abnormalities2. Teratogenesis
due to Drugs: The potential danger of new drugs to the fetuses was exemplified by two
unrelated drugs, thalidomide and meclizine. Thalidomide is a mild sedative tranquilizer that was
prescribed for use in pregnancy in many European countries in the late 1950s. The German
scientist Lenz (1961) reported a possible connection between this drug and an increased
frequency of a human congenital abnormality of the limbs known as Amelia (no limbs) and the
closely related phocomelia (no development of long bones of limbs and flipperlike hands or feet
attached directly to the trunk). A daily intake of thalidomide for one week during early pregnancy
was sufficient to induce limb defect. From 1959-1961 thousands of babies in West Germany
and hundreds in other countries such as Japan, were born with partial or complete absence of
limbs or limbs with defects. This has led to extreme caution in the introduction of new drugs for
commercial distribution, for pregnant mothers, since they may cause irreparable harm to human
embryos. Antimitotic drugs used in cancer therapy would be especially harmful to the rapidly
growing embryo. However, only aminopterin (a folic acid antagonist) has proved to be
teratogenic in man. This chemical has been used to bring about abortion. When it fails to induce
abortion, the offspring is likely to show multiple malformations. Teratogenic effects of certain
other drugs such as quinine (for malaria), busulphan (for leukemia), and chlorambucil (for
Hodgkin’s disease) have also been reported. 3. Teratogenesis due to radiation: It is
recognized that the use of roentgen rays or radium in the treatment of pregnant patients having
pelvic tumors, is possible to produce skeletal abnormalities in the fetuses. The contact of
pregnant women with severe atomic radiation, as in the Hiroshima and Nagasaki explosions, led
to a fetal death rate of about 40 percent. The infants who survived tended to show confirmation
of brain-cell damage. 4. Teratogenesis due to autoimmunization: Significant interest was
aroused by the examination that mothers of infants born without thyroid glands, i.e., thyroid
cretins, may have antithyroid antibodies in their blood. This observation suggests that products
of embryonic organ. primitive may cross the placenta, inducing s maternal antibody production
the antibodies may return to embryo and interfere with organ differentiation 5. Teratogenesis
due to malnutrition: It is a well-known fact that deficiency of various vitamins causes several
diseases in man the deficiency of vitamin A causes night blindness, the deficiency of vitamin D
leads to abnormal development of bone and teeth and the deficiency of vitamin K causes
abnormalities in the clotting of blood. It has been studied that if rats, rabbits, and other animals
are given a diet deficient in vitamin A, B, and D, they may produce various teratogenic effects
such as hare- lip, cleft palate, spina bifida, skeletal defects, brain defects, etc. Likely, such
vitamin malnutrition in pregnant mothers of the human species may also cause similar
congenital malformation.

Sensitive period of the teratogen: The developmental stage at which the embryo is treated by
a teratogen has great relevance to teratogenesis. In general, the very early stages of
development are not much affected by the teratogen, possibly due to the considerable
regulatory capacity of the embryo. Similarly, the later stages of development, when maturation
and growth of organs occur, are also comparatively resistant to teratogens. The main
teratogenic period starts with the creation of germ layers and continues up to organogenesis.
For example, in rats trypan blue (20 mg intravenous injection to the mother) and actinomycin D
(0.3 mg/kg body weight to the mother) are maximally teratogenic on the 8th and 9th day of
gestation, respectively. Both chemicals are ineffective after 10th day and only minimally
teratogenic before the 6th day of gestation.
Specificity of the teratogen: Almost any teratogen can produce almost any terata if applied at
the right time in the experimental animal. However, each teratogen produces its typical
syndrome. For example, the micromelia (short limb) in fowl can be caused by a deficient or
imbalanced diet, (i, e., riboflavin (or biotin deficiency), insulin, thallium, boric acid, pilocarpine,
propanediol-1, 3 sulphanilamides or serine sulfate. A similar situation occurs in the case of
tetraptera (four wings) in Drosophila melanogaster, i.e., phenocopies of tetraptera can be
obtained by treating the early embryo with either ether or 40°C heat shock. However, the types
of micromelia produced in fowl by different teratogens are only superficially similar. The type of
malformation produced by a teratogen is also dependent on the developmental stage of an
embryo at the time of treatment. For example, insulin (2 units in yolk sac), causesrumplessness
in the young fowl embryo (24 hours) but micromelia and abnormal beak in the older embryos
(70 to 170 hours) of fowl. Thus, each organ system appears to have its sensitive period for a
given teratogen.

Teratogenic dose: A teratogen may be quite ineffective at a very low dose, while at its high
doses it may be lethal to all embryos. A teratogenic dose range comprises doses that produce
at least some malformed and living embryos. It is also necessary to discriminate between the
teratogenic and the toxic effects of an agent. By definition, an agent is toxic, if it causes
embryonic death by direct action and not through teratogenic interaction. For example, trypan
blue (50 mg/kg) injected into 8.5-day pregnant rats affects 65 percent of the embryos. But with
advancing gestation, out of these 65 percent of the embryos the number of dead one increases
at the expense of live and malformed individuals. Moreover, when trypan blue is injected outside
the teratogenic period, it does not result in appreciable [Link] blue therefore is a
teratogenic and not toxic [Link] of Teratogens with other Environmental
Factors: The effect of any teratogen depends on other factors present in the environment of an
organism. Many types of communication between environmental factors do occur. For example,
the teratogenic effects of insulin on a 96 120- hour-old fowl embryo are almost completely by
the simultaneous injection of nicotinamide. The synergistic effects of agents are also well
known. The incidence of both the micromelia and abnormal beak in the insulin-injected fowl
embryos is significantly enhanced by chlorpromazine, a non- teratogenic but slightly toxic
chemical. Likewise in another type of interaction the nonteratogenic dos of two teratogens,
sulphanilamide and 6-aminonicotinamide, can cause teratogenesis when injected altogether.
Developmental mechanisms of teratogenesis: Each phase of development is prone to a
defect. The chronological nature of development is based on networks of interacting systems,
what happens at one stage is essential to all consequent stages. Defects initiated at an early
stage may be expressed at later stages because intermediate steps are abnormal. The
magnitude of a defect will depend on the stage of development at which it originates and the
development process that is a specific target. Lesions arising at early stages usually have more
cells are affect (Grant, 1978). Since in terata any organ and any tissue may be affected so the
mode of production of terata is much varied. The development processes that may be affected
by teratogenesis include competence, induction (evocation), determination, histogenesis and
morphogenesis, cell growth, cell division, cell death, and cell locomotion. The genetic and
molecular studies on development indicate that each teratogen affects the developmental
processes by essentially affecting cell metabolism and gene expression. This is brought about
by several independent or inter-related mechanisms such as: 1) Production of defective,
deficient, or no proteins 2) Production of proteins at an inappropriate developmental juncture 3)
Production of defective, deficient, or no rRNA or tRNA of a particular kind or 4) Alteration in
membrane permeability. For example, in the case of sickle cell anemia, the production of a
defective protein (β chain of hemoglobin) leads to sickle cell syndrome.

NOTE:ALSO ADD TYPES DESIGNATED BY PROFESSORS

 Metaplasia is a reversible cellular adaptation in which one mature, differentiated cell type is
replaced by another mature, differentiated cell type. The new cell type is better equipped to
withstand the environmental stress or chronic irritation that caused the change.

Key characteristics:Reversible: If the stressor is removed, the metaplastic tissue can return to
its normal cell [Link]: The change is an adaptive response to environmental stress or
[Link] from precursor cells: The transformation occurs through the reprogramming of
local tissue stem cells or undifferentiated mesenchymal cells.

Mechanism:The change in cell type is driven by the reprogramming of precursor cells (stem
cells) found in the affected [Link]: Chronic irritation or inflammation triggers a response
mediated by cytokines, growth factors, and components of the extracellular
[Link]: These signals cause the precursor cells to differentiate along a new
pathway, producing a different, more resilient cell [Link]: The new cells form a
mature tissue that is different from the original, even though they originated from the same
precursor population.
Types of metaplasia
Metaplasia is broadly classified into two main types: epithelial and mesenchymal.

1. Epithelial metaplasia:This is the most common form, involving the conversion of one type
of epithelial cell to another.
 Squamous metaplasia:Description: A type of epithelial metaplasia where columnar epithelium
is replaced by stratified squamous [Link]: Squamous cells are hardier and can
better withstand physical stress. However, this change can lead to a loss of specialized function,
such as mucus secretion and ciliary [Link]:Respiratory tract: In chronic
smokers, the normal ciliated pseudostratified columnar epithelium of the bronchi is replaced by
stratified squamous [Link] cervix: The simple columnar epithelium of the
endocervix is replaced by squamous epithelium, which can be triggered by human
papillomavirus (HPV) infection or chronic [Link] bladder: Chronic infection or bladder
stones can cause the transitional epithelium to change into squamous epithelium.

 Columnar metaplasia:Description: The replacement of squamous epithelium with columnar

[Link]:Barrett's esophagus: In gastroesophageal reflux disease (GERD),
chronic acid exposure causes the stratified squamous epithelium of the lower esophagus to be
replaced by specialized intestinal-type columnar epithelium with goblet cells. This is an adaptive
change, as the columnar cells are better able to secrete protective [Link] intestinal
metaplasia: Chronic inflammation in the stomach, often due to Helicobacter pylori infection, can
lead to the replacement of gastric mucosa by intestinal-type epithelium.

2. Mesenchymal metaplasia:This involves the conversion of one type of connective tissue into
another.
 Osseous metaplasia:Description: The formation of bone within connective tissues, such as
fibrous tissue, cartilage, or [Link]:Myositis ossificans: Following muscle trauma or
inflammation, muscle tissue is replaced by [Link] of fractures: The formation of a
cartilaginous callus, which later ossifies, is part of the normal healing process.

 Cartilaginous metaplasia:Description: The formation of cartilage in an abnormal

[Link]: Sometimes seen in the connective tissue of elderly people.
 Clinical significance:Metaplasia is a significant finding that requires attention, though it is not
cancer. Loss of function: While the new cell type is better at resisting stress, it loses the
specialized functions of the original tissue. For example, squamous metaplasia in the respiratory
tract results in the loss of mucus production and ciliary clearance, increasing the risk of
[Link] condition: Persistent exposure to the damaging stimulus can cause
metaplasia to progress to dysplasia (disordered, atypical cell growth) and eventually to
malignant neoplasia (cancer).Reversibility and treatment: The best course of action is to
identify and remove the underlying cause. For example, smoking cessation can reverse
respiratory metaplasia, and treating acid reflux can improve Barrett's esophagus. However,
some changes, like intestinal metaplasia, may be permanent once established, even with
[Link]: Areas of metaplasia, especially those linked to precancerous risk, must
be monitored regularly to detect any progression to dysplasia.
Vitellogenesis is the process of yolk formation and accumulation in an oocyte, providing
nutrients for embryonic development. It involves the synthesis of precursor proteins
called vitellogenins outside the oocyte (often in the liver of the female parent), which are then
transported to the oocyte via the circulatory system. Specific receptors on the oocyte membrane
mediate the uptake of these vitellogenins, which are converted into yolk proteins (vitellins) and
stored within the oocyte. This process is hormonally regulated and essential for the survival of
the developing embryo in oviparous (egg-laying) animals like birds, reptiles, fish, and
amphibians. Vitellogenesis is the process of yolk formation in oviparous animals, where nutrient-
rich yolk is synthesized and deposited into the oocyte (egg cell). This process involves the
production of vitellogenins (yolk precursor proteins) under hormonal control,
primarily estrogens, and their uptake into the oocyte to provide nourishment for the developing
embryo. The key stages include pre-vitellogenesis (oocyte growth), vitellogenesis (yolk
deposition), and the utilization of the deposited yolk during embryogenesis.

Key Aspects of Vitellogenesis:Yolk Formation: Vitellogenesis is essentially the synthesis

and accumulation of yolk, a vital nutrient store for the [Link]: These are the
main protein precursors of yolk and are synthesized by the liver or fat body in response to
hormones. Hormonal Control: Hormones, particularly estrogens, are crucial for inducing
vitellogenesis by stimulating vitellogenin production in the liver or fat body.

 Oocyte Uptake: Once synthesized, vitellogenins circulate in the hemolymph (blood) and are
taken up by the oocyte through endocytosis, where they are then processed into the mature
yolk. Stages
1. Pre-vitellogenesis:
This initial phase involves oocyte growth and preparation for yolk deposition, characterized by
high nucleolar activity and the increased production of cytoplasmic substances like
ribonucleoproteins and mitochondria.
[Link]: This is the active phase of yolk accumulation within the oocyte, where
vitellogenins are internalized and organized into yolk granules or spheres.
Hormonal Regulation

 Estrogens: Circulating estrogens, produced by the ovary from maturing follicles, are the
primary stimulus for vitellogenin synthesis.
 Juvenile Hormone: In many insects, juvenile hormone, secreted by the corpora allata,
stimulates the fat body to produce vitellogenin and promotes oocyte growth.
 Other Hormones: Other hormones, such as progesterone, and possibly ovarian GnRH and
bradykinin, can also influence vitellogenesis.
Significance

 Nutritional Support: Yolk provides essential nutrients for the developing embryo, influencing
its growth and differentiation.
 Developmental Timing: The timing of vitellogenesis is synchronized with other physiological
processes like feeding and mating to ensure successful reproduction.
 Egg Size and Development: The amount of yolk deposited directly influences egg size and
can affect cell division patterns, morphogenetic movements, overall developmental maturity.
The growth phase of oocyte is divided into two stages, namely previtellogenesis and
vitellogenesis i. Previtellogenesis During previtellogenesis the cytoplasm and nuclear
materials of primary oocyte grow and increase considerably in volume. The yolk and other
food materials are not synthesized during this phase. The following changes occur during
previtellogenesis The nuclear sap is produced in large amount. As a result, the nucleus
increases in size. The large nucleus of the oocyte is called germinal vesicle Homologous
chromosomes pair together In amphibians, the chromosomes of primary oocytes acquire a
characteristic shape; thin loops or threads appear on the sides of the chromosomes. These
loops give a brush-like appearance to the chromosomes. Hence the chromosomes are called
lamp-brush chromosomes. The loops of these chromosomes are actively involved in the
synthesis of mRNA The ribosomal RNAs are produced in a remarkable amount. They are
produced by the nucleolus. As a result the nucleolus increases greatly in size. The genes
producing the rDNA are multiplied several times to facilitate the rapid synthesis of rDNA. This
increase in the number of genes is called amplification In many cases, the production of RNA is
increased further, by the development of a greater number of nucleoli (a small dense spherical
structure in the nucleus of a cell during interphase). This phenomenon is more common in the
egg of the amphibian. The mitochondria increase in number Cortical granules are
manufactured by the cistemae of Golgi complex The growing oocytes are surrounded by
special kinds of nutritive cells. These cells immensely assist the growth of oocytes in various
ways. There are two types of nutritive cells, namely follicle cells and nurse cells In the ovary
of chordates, the developing oocyte is surrounded by follicle cells In mammals, the follicle
cells and the developing ovum together constitute a Graafian follicle. Each Graafian follicle
consists of a cavity called antrum filled with a fluid called liquor folliculi. The cavity is
surrounded by three layers, namely an outer theca externa, a middle theca interna and an
inner membrana granulosa. The oocyte lies inside the antrum. It is surrounded by a few
layers of follicle cells called corona radiata. The oocyte is attached to the membrana
granulosa on the side by a stalk of cells called discus proligerus. The oocyte is surrounded by
a transparent membrane called zona pellucid.

[Link] The process of formation and deposition of yolk in the oocyte is called
vitellogenesis. Yolk is the nutritive material of the ovum. It is present in the form of platelets
or granules Origin of Yolk Initially when an oocyte starts developing, it does not contain any
nutrients. The nutrients are formed only during growth phase of oogenesis. There are two
views regarding the place of the origin of yolk Insitu Origin: A very small amount of yolk is
synthesized in the oocyte cytoplasm. For example, in vertebrates less than 1% is synthesized
by the oocyte cytoplasm Exogenous Origin: A major part of the yolk is synthesized outside the
oocyte. In vertebrates, it is synthesized in the liver; in insects it is synthesized in the fat body
Transportation of Yolk In vertebrates, the yolk synthesized in the liver, is in a soluble state. It
is transported by the blood stream to the follicle cells present around the oocyte. The follicle
cells deposit the yolk in the oocyte. The transport of yolk to the oocyte is facilitated by the
development of finger-like structures called microvilli by the oocyte and macrovilli by the follicle
cells The deposit of yolk and other nutrients like the lipid, glycogen, etc. begins close to the
oocyte surface and fills the peripheral cytoplasm first. Gradually, the entire cytoplasm is
deposited with yolk except the perinuclear zone. Thus the cytoplasm is progressively occupied
from the peripheral zone inward.