Unit 2: Early Embryonic Development

Embryonic induction and organizers

 Embryonic induction
• During embryogenesis of multicellular organisms, some cells are induced to
differentiate into specific organs or tissues by the presence of other cells. This
process of inducement by cells is called embryonic induction or dependent
differentiation.
• The embryonic induction is a morphogenetic effect in which one embryonic
tissue transmits a chemical stimulus that influences another embryonic part
to produce a structure that otherwise would not come into existence.
• The embryonic tissue that exerts the inductive influence is known as
inductor.
• The responder, is the cell or tissue being induced. Cells of the responding
tissue must have both a receptor protein for the inducing factor and the
ability to respond to the signal.
• The ability to respond to a specific inductive signal is called competence
(Waddington 1940).
 Embryonic induction
Organizer
The effect of embryonic interaction or organizer is a morphogenetic effect by
which one organic tissue transmits a chemical substance that influences other
embryonic part to produce a structure that otherwise could not come into
existence. The embryonic tissue which exerts such an influence is called an
inductor and the chemical substance secreted by an inductor is known as
evocators. The tissue on which evocator works and the tissue responses is
known as responsive tissue. The action of the indicator through evocator is
known as induction action or organizer action. This process of induction
influences greatly the protein synthesis mechanism of responsive tissues as a
result of which definite structure forming cells become very active.
 Embryonic induction
Origin and concept of Organizer
 Embryonic induction
Primary organizer:
Spemann continued his grafting experiments taking tissues from different zones of the
gastrula and observed that except dorsal lip of the early gastrula other zone of tissue
can not create any induction effect but when dorsal lip is grafted a complete embryo is
formed. He named the dorsal lip as organizer as this dorsal lip organizes the
developmental process of the embryo. According to him this dorsal lip induces to form
neural tube and the neural tube then induces to form the eyes. The dorsal lip is
composed of chorda-mesoderm and as it primarily acts as inducer so he named the
dorsal lip or chordamesoderm as primary organizers.

Secondary, tertirary and quaternary organizers

As the gastrulation proceeds due to primary organizer’s induction primary organs begin
to form and the early stages of organ development are known as organ rudiments.
These organ rudiments themselves may act as organizer and then they are known as
secondary organizer. Tissues formed by the action of secondary organizer may in turn
induce further development. Then they are known as tertiary organizer. These
successive stages of organizer activities start from the primary organizer.
 Embryonic induction
Inductive interactions
Howard Holtzer (1968) distinguished two major modes of inductive interaction.
1) Instructive interaction
• In instructive interaction, a signal from the inducing cell is necessary for initiating new
gene expression in the responding cell. Without the inducing cell, the responding cell
is not capable of differentiating in that particular way.
• For example, one instructive interaction is when a Xenopus optic vesicle
experimentally placed under a new region of head ectoderm causes that region of the
ectoderm to form a lens
2) permissive interaction
• The second type of inductive interaction is permissive interaction. Here, the
responding tissue has already been specified and needs only an environment that
allows the expression of these traits.
• For instance, many tissues need an extracellular matrix to develop. The extracellular
matrix does not alter the type of cell that is produced, but it enables what has already
been determined to be expressed.
 Embryonic induction
1) Instructive interactions
Ectodermal competence and the
ability to respond to the optic
vesicle inducer in Xenopus.
• The optic vesicle is able to induce
lens formation in the anterior
portion of the ectoderm (1)
• but not in the presumptive trunk
and abdomen (2).
• If the optic vesicle is removed (3),
the surface ectoderm forms
either an abnormal lens or no
lens at all.
• Most other tissues are not able to
substitute for the optic vesicle (4).
 Embryonic induction
Inductive interactions
2) permissive interaction
A dramatic example of permissive interactions at
work comes from the regenerative medicine field, in
which an extracellular matrix scaffold can promote
the differentiation and rebuilding of a beating heart.
Doris Taylor’s research group used detergents to
remove all the cells from a cadaveric rat heart, which
leaves behind the natural extracellular matrix (Ott et
al. 2008). Proteins like fibronectin, collagen, and
laminin held together the rest of the ECM and
maintained the intricate shape of the heart. The
researchers then infused this ECM scaffold with
cardiomyocytes. Surprisingly, these cells
differentiated and organized into a functionally
contracting “recellularized” heart. Therefore, the
environmental conditions of the decellularized ECM
were permissive in allowing the cardiomyocytes to
recreate contracting heart muscle.
 Embryonic induction
Epithelial-mesenchymal interactions:
Regional specificity of induction
The chick epidermis secretes proteins that signal the underlying dermal cells to
form condensations, and the condensed dermal mesenchyme responds by
secreting factors that cause the epidermis to form regionally specific cutaneous
structures (Nohno et al. 1995; TingBerreth and Chuong 1996). These structures
can be the broad feathers of the wing, the narrow feathers of the thigh, or the
scales and claws of the feet. The dermal mesenchyme is responsible for the
regional specificity of induction in the competent epidermal epithelium.
 Embryonic induction
Regional specificity of induction
Researchers can separate the embryonic epithelium and mesenchyme from each
other and recombine them in different ways (Saunders et al. 1957). The same
epithelium develops cutaneous structures according to the region from which
the mesenchyme was taken. Here, the mesenchyme plays an instructive role,
calling into play different sets of genes in the responding epithelial cells.
 Embryonic induction
Epithelial-mesenchymal interactions:
Genetic specificity of induction
Whereas the mesenchyme may instruct the epithelium as to what sets of genes
to activate, the responding epithelium can comply with these instructions only so
far as its genome permits.
This property was discovered through experiments involving the transplantation
of tissues from one species to another.
 Embryonic induction
Epithelial-mesenchymal interactions:
Genetic specificity of induction
Hans Spemann and Oscar Schotté (1932) placed
presumptive flank ectodermal tissue from a newt
gastrula into the presumptive oral regions of frog
embryos. The structures of the mouth region differ
greatly between salamander and frog larvae. The
salamander larva has club-shaped balancers beneath
its mouth, whereas the frog tadpole produces mucus-
secreting glands and suckers. The frog tadpole also has
a horny jaw without teeth, whereas the salamander
has a set of calcareous teeth in its jaw. The larvae
resulting from the transplants were chimeras. The
salamander larvae had frog-like mouths, and the frog
tadpoles had salamander teeth and balancers.
In other words, the mesenchymal cells instructed the
ectoderm to make a mouth, but the ectoderm
responded by making the only kind of mouth it “knew”
how to make, no matter how inappropriate
 Embryonic induction
Inducer molecules
• When membrane proteins on one cell surface interact with receptor proteins
on adjacent cell surfaces (as seen with cadherins), the event is called a
juxtacrine interaction (since the cell membranes are juxtaposed).
• Paracrine factors are proteins made by a cell or a group of cells that alter the
behavior or differentiation of adjacent cells. In contrast to endocrine factors
(hormones), which travel through the blood and exert their effects on cells
and tissues far away, paracrine factors are secreted into the extracellular
space and influence their close neighbors.
• Autocrine interactions occur when the same cells that secrete the paracrine
factors also respond to them.
 Embryonic induction
Classification of induction:
Lovtrup (1974) classified induction into two principal classes:
a) Endogenous induction: Shapes and sizes of some of the embryonic cells
changes after secreting inducting substances and this induction brings
about differentiation of cells. As for example small cells of the dorsal lip
carrying yolk granules act as endogenous induction.
b) Exogenous induction: When either by external influence or by contact any
cell or tissue induces nearby tissue to differentiate, then it is known as
exogenous induction.
Exogenous induction may again be of two types. As-
1. Homotypic: When the contact induction induces to form same types
of cells, it is known as homotypic.
2. Heterotypic: When the contact induction induces different types of
cell differentiation, it is known as heterotypic induction.
 Embryonic induction
Types of inductors:
On the basis of the regional specificity, Lehmon (1945) classified the inductors
into three groups. They are:
1) Archenocephalic inductor: Due to induction effect of this inductor partial
head, fore-brain, eye, nasal cavities are formed.
2) Deuterencephalic inductor: By its induction effect posterior portion of the
head, ear cavities etc. are formed.
As arechenocephalic and deuterencephalic inductors induce the formation of
different parts in the head region so they together are known as cephalic or
head inductors.
3) Spino-caudal inductor: Their inductive influence leads to the formation of
spinal cord and different structures of the tail region
Unit 2: Early Embryonic Development

Gastrulation in Frog
• an embryo is divided into two hemispheres: the animal pole and
the vegetal pole within a blastula. The animal pole consists of small cells that
divide rapidly, in contrast with the vegetal pole below it. ... The animal
hemisphere is dark brown, and the vegetal hemisphere is only weakly
pigmented.
• In frogs, the egg cell is a massive cell—much larger than a normal frog cell—
and it has an uneven distribution of various molecules, which are deposited in
the egg by the mother frog before fertilization. This asymmetry is even visible
in the egg: it has a dark-colored top—called the animal pole—and a light, yolky
bottom—called the vegetal pole. Many mRNA and proteins from the mother
frog are distributed unevenly between the animal and vegetal poles.
 Frog Male gamete
• Mature spermatozoa of a frog is made up of 3
parts i.e., head, middle piece and tail.
• Middle piece region is very small as compared to
that of the other sperms,
• The head consists of acrosome and large nucleus.
The acrosome secretes enzymes which helps in
penetration into the egg.
• The middle piece of spermatozoon is composed of
almost entirely of mitochondria and centriole
(Centriole helps in the spindle formation during
cleavage).The mitochondrial apparatus supplies
the required amount of energy in the form of ATP.
• The tail of sperm is flagellar and very long. It helps
in locomotion and thus is very essential for the
mobility of the sperm.
 Frog gametes

Male gamete Female gamete

Amplexus Newly laid clutch of Eggs
The brown area of each egg is the pigmented
animal hemisphere. The white spot in the
middle of the pigment is mwhere the egg's
nucleus resides
 Fertilization
• Fertilization can occur anywhere in the animal hemisphere of the amphibian embryo.
• When the sperm enters the egg, the cytoplasm at the edge of the egg cell, called the cortical cytoplasm,
rotates 30 degrees towards the site of sperm entry.
• Rotation exposes a wedge of the cytoplasm underneath, sometimes producing a visible zone of lighter
color called the gray crescent.
• The point of sperm entry is important because it determines the orientation of the dorsal-ventral axis of
the larva (tadpole). The point of sperm entry will mark the ventral side of the embryo, while the site 180
degrees opposite the point of sperm entry will mark the dorsal side.
• the site opposite the point of sperm entry will be the place where gastrulation begins, and the
microtubular array will become extremely important in initiating the dorsal-ventral and anterior-
posterior axes of the larva.
 Unequal radial holoblastic cleavage
• Cleavage in most frog and salamander
embryos is radially symmetrical and
holoblastic
• The amphibian egg, however, contains much
more yolk. This yolk, which is concentrated in
the vegetal hemisphere, is an impediment to
cleavage. Thus, the first division begins at the
animal pole and slowly extends down into the
vegetal region
• In many species (especially salamanders and
frogs of the genus Rana), the first cleavage
division bisects the gray crescent.
 Unequal radial holoblastic cleavage
• While the cleavage furrow is still cleaving the yolky cytoplasm of the vegetal
hemisphere, the second cleavage has already started near the animal pole.
This cleavage is at right angles to the first one and is also meridional
• The third cleavage is equatorial. However, because of the vegetally placed
yolk, the third cleavage furrow is not actually at the equator but is displaced
toward the animal pole. It divides the amphibian embryo into four small
animal blastomeres (micromeres) and four large blastomere, (macromeres)
in the vegetal region
 Unequal radial holoblastic cleavage
• Despite their unequal sizes, the blastomeres continue to divide at the same rate
until the twelfth cell cycle (with only a small delay of the vegetal cleavages). As
cleavage progresses, the animal region becomes packed with numerous small cells,
while the vegetal region contains a relatively small number of large, yolk-laden
macromeres
• An amphibian embryo containing 16 to 64 cells is commonly called a morula (plural
morulae; Latin, "mulberry," whose shape it vaguely resembles). At the 128-cell
stage, the blastocoel becomes apparent, and the embryo is considered a blastula
 Unequal radial holoblastic cleavage
• The amphibian blastocoel serves two major
functions.
i) First, it permits cell migration during
gastrulation; and
ii) second, it prevents the cells beneath it
from interacting prematurely with the
cells above it.
• Numerous cell adhesion molecules keep the
cleaving blastomeres together. One of the
most important of these is EP-cadherin.
 The mid-blastula transition: Preparing for gastrulation
• An important precondition for gastrulation is
the activation of the genome.
• At that time, the embryo experiences a mid-
blastula transition (MBT) different genes
begin to be transcribed in different cells, the
cell cycle acquires gap phases, and the
blastomeres acquire the capacity to become
motile.
 Amphibian Gastrulation
Vegetal rotation and the invagination of the bottle cells
• Gastrulation initiates at the future dorsal side of the embryo, just below the equator, in
the grey crescent region.
• Vegetal rotation places the prospective pharyngeal endoderm cells adjacent to the
blastocoel and immediately above the involuting mesoderm. These cells then migrate
along the basal surface of the blastocoel roof, traveling toward the future anterior of the
embryo
• The superficial layer of marginal cells is pulled inward to form the endodermal lining of
the archenteron, merely because it is attached to the actively migrating deep cells
 Amphibian Gastrulation
Involution at the blastopore lip
• The next phase of gastrulation involves the involution of the marginal zone cells while the
animal cells undergo epiboly and converge at the blastopore
• When the migrating marginal cells reach (and become) the dorsal lip of the blastopore, they
turn inward and travel along the inner surface of the outer animal hemisphere cells (i.e., the
blastocoel roof).
• The first cells to compose the dorsal blastopore lip and enter into the embryo are the
prospective pharyngeal endoderm of the foregut (including the bottle cells). As these first cells
pass into the interior of the embryo, the dorsal blastopore lip becomes composed of cells that
involute into the embryo to become the prechordal plate (the precursor of the head
mesoderm).
 Amphibian Gastrulation
Involution at the blastopore lip
• The next cells involuting into the embryo through the dorsal blastopore lip are the
chordamesoderm cells. These cells will form the notochord, a transient mesodermal rod that
plays an important role in inducing and patterning the nervous system.
• As the new cells enter the embryo, the blastocoel is displaced to the side opposite the dorsal
lip of the blastopore. Meanwhile, the lip expands laterally and ventrally as the processes of
bottle cell formation and involution continue around the blastopore.
• The widening blastopore "crescent’ develops lateral lips and finally a ventral lip over which
additional mesodermal and endodermal precursor cells pass.
 Amphibian Gastrulation
Involution at the blastopore lip
• With the formation of the ventral lip, the blastopore has formed a ring
around the large endodermal cells that remain exposed on the vegetal
surface
• This remaining patch of endoderm is called the yolk plug; it, too, is
eventually internalized. At that point, all the endodermal precursors
have been brought into the interior of the embryo, the ectoderm has
encircled the surface, and the mesoderm has been brought between
them.
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• involution begins dorsally, led by the pharyngeal endoderm and the
head mesoderm. These tissues will migrate most anteriorly beneath
the surface ectoderm.
• Meanwhile, as the lip of the blastopore expands to have dorsolateral,
lateral, and ventral sides, the prospective heart, kidney, and ventral
mesodermal precursor cells enter into the embryo
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• The IMZ(intermediate marginal zone) is originally several layers thick. Shortly before their
involution through the blastopore lip, the several layers of deep IMZ cells intercalate radially to
form one thin, broad layer.
• This intercalation further extends the IMZ vegetally
• At the same time, the superficial cells spread out by dividing and flattening. When the deep
cells reach the blastopore lip, they involute into the embryo and initiate a second type of
intercalation. This intercalation causes a convergent extension along the mediolateral axis that
integrates several mesodermal streams to form a long, narrow band
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• the mesodermal stream continues to migrate toward the animal pole, and the overlying layer
of superficial cells (including the bottle cells) is passively pulled toward the animal pole,
thereby forming the endodermal roof of tile archenteron
• The radial and mediolateral intercalations of the deep layer of cells appear to be responsible
for the continued movement of mesoderm into the embryo.
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• As mesodermal movement progresses, convergent extension continues to narrow and
lengthen the involuting marginal zone.
• The involuting cells contain the prospective endodermal roof of the archenteron in its
superficial layer and the prospective mesodermal cells, including those of the notochord, in its
deep region
• Toward the end of gastrulation, the centrally located notochord separates from the somitic
mesoderm on either side of it, and the notochord elongates separately as its cells continue to
intercalate
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• The dorsal portion of the noninvoluting marginal zone (NIMZ) extends more rapidly toward
the blastopore than the ventral portion does, causing the blastopore lips to move toward the
ventral side. While those mesodermal cells entering through the dorsal lip of the blastopore
give rise to the central dorsal mesoderm (notochord and somites), the remainder of the body
mesoderm (which forms the heart, kidneys, bones, and parts of several other organs) enters
through the ventral and lateral blastopore lips to create the mesodermal mantle.
 Amphibian Gastrulation
CONVERGENT EXTENSION OF THE DORSAL MESODERM
• The endoderm is derived from the superficial cells of the involuting marginal zone that form
the lining of the archenteron roof and from the subblastoporal vegetal cells that become the
archenteron floor. The remnant of the blastopore-where the endoderm meets the ectoderm-
now becomes the anus.
 Amphibian Gastrulation
Epiboly of the prospective ectoderm
• During gastrulation, the animal cap and noninvoluting marginal zone cells expand by epiboly
to cover the entire embryo.
• These cells will form the surface ectoderm
Amphibian Gastrulation