Reproduction and Development

Zelmay Jan
July 2023

Table of Contents
1 Introduction 3

2 Asexual and Sexual Reproduction 3

2.1 Asexual Reproduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.2 Variations in Sexual Reproduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.3 Reproductive Cycles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

3 Fertilization 5

4 Development 6
4.1 General Development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.2 Accessory Sex Organs and External Genitalia Development . . . . . . . . . . . . . . 7

5 Endocrine Regulation 8

6 Male Reproductive System 8

6.1 Anatomy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
6.2 Gonadotropin Secretion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
6.3 Spermatogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
6.4 Accessory Sex Organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
6.5 Erection, Emission, and Ejaculation . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

7 Female Reproductive System 13

7.1 Anatomy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
7.2 Ovarian Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
7.3 Menstrual Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
7.4 Changes in the Endometrium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
7.5 Menopause . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

8 Fertilization, Lactation, and Birth 19

8.1 Capacitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
8.2 Fertilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
8.3 Birth and Pregnancy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
8.4 Lactation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

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9 Disorders 23
9.1 Hermaphroditism and Pseudohermaphroditism . . . . . . . . . . . . . . . . . . . . . 23
9.2 Endometriosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
9.3 Ectopic Pregnancy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
9.4 Genetic Screening . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

10 Contraceptives 24
10.1 Rhythm Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
10.2 Coitus Interrupts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
10.3 Oral Contraceptives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
10.4 Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
10.5 Abortion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

11 Development 26
11.1 Cleavage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
11.2 Gastrulation in Frogs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
11.3 Gastrulation in Chicks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
11.4 Gastrulation in Humans . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
11.5 Organogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
11.6 Cytoskeleton in Morphogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
11.7 Fate Mapping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
11.8 Axis Formation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
11.9 Induction in Pattern Formation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

12 Conclusion 32

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1 Introduction
We would like to issue a gentle warning that the content discussed within this handout may be
sensitive and explicit. Reproduction is a fundamental biological process that allows living organ-
isms to perpetuate their species, but it involves topics that might be uncomfortable or triggering
for some individuals.

Welcome to our guide on reproduction and development, a fundamental mechanism that per-
petuates life across all forms of organisms inhabiting our planet. In the following pages, we delve
into the intricacies of this biological phenomenon, shedding light on its diverse processes and
underlying principles.

2 Asexual and Sexual Reproduction

2.1 Asexual Reproduction
Asexual reproduction is a form of reproduction that does not involve fertilization; it usually relies
entirely on mitotic cell division and produces a genetically identical individual. This simple form
of reproduction is generally exclusive to invertebrates.
• In budding, a section of the child forms from an outgrowth of the parent organism. However,
the outgrowth remains attached to the parent organism. This form of reproduction occurs
in stony corals.

• Fission splits the parent organism into two individuals of even size.

• In fragmentation, part of the parent organism breaks off. The lost body parts that form
from fragmentation then undergo regeneration. This mechanism is found in annelids,
corals, cnidarians, and tunicates.

Figure 1: Fragmentation in Planarians. (Source: funscience)

• In parthenogenesis, the egg develops without being fertilized. This occurs in bees, wasps,
and ants.

– In bees, unfertilized queen eggs form haploid males via parthenogenesis.

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• Parthenogenesis is also known to occur in vertebrates in response to low population density.

Komodo dragons, hammerhead sharks, and even sawfish have been observed to undergo
parthenogenesis.

2.2 Variations in Sexual Reproduction

Sexual reproduction, in most animals, requires the mating of a male and female. The rest of the
handout will be dedicated to describing this process in humans. However, in many animals, finding
a partner for reproduction is difficult; animals solve this problem by removing the distinction
between the two genders, creating a hermaphrodite, for which the organism has reproductive
systems of both genders.

• In hermaphrodites, which include corals and sea slugs among many other species, any two
individuals can mate since they both have male and female reproductive systems. In certain
corals, self-fertilization can also occur.

• In the blue-headed Wrasse, males live in a harem with several females. When the lone male
dies, the biggest female undergoes sex reversal to replace the male. This form of reproduction
is known as protogyny (gyno = female, proto = first).

• Oysters, much like the blue-headed wrasse, undergo sex reversal. However, oysters begin as
males and transition to females as they gain size. This mechanism is called protandry.

– This strategy is favored due to the positive correlation between a female’s size and the
number of gametes she produces. In other words, it is more favorable to be female when
bigger.

2.3 Reproductive Cycles

Most animals display cycles in their reproductive activity. This way, animals can produce offspring
only when their chances of survival are optimal.

• In some animals, climate change has resulted in a large reduction in reproductive success,
such as in caribou.

– Caribou migrate to birthing grounds in the spring to eat and to birth calves. They
determine the time of their visit based on the length of the day in order to optimize
the amount of grass available when they visit. As a result of climate change, there is a
mismatch between the length of day and the time of grass sprouting, resulting in less
food available to caribou and thus a decline in their population.

• Reproductive cycles can also be linked to asexual reproduction. In Daphnia, for example,
asexual reproduction (i.e. parthenogenesis) occurs during times of adverse environmental
conditions.

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• Aspidoscelis is a genus of asexual whiptail lizards that

still observe mating rituals despite being fully asex-
ual.

– When estrogen levels are high in an individual, it

mimics female mating patterns. However, if pro-
gesterone levels are high, it mimics male mating
patterns. One female of each mating pair has to
mimic a male.

This leads to an interesting discussion on the evolutionary

background behind sexual reproduction. One may argue
that asexual reproduction is superior when considering the
number of offspring an individual can have compared to a
sexually reproducing individual. However, sexual reproduc-
tion allows for genetic diversity and thus greater adaptation
compared to asexual reproduction. Thus, it can be gener-
alized that asexual reproduction is more valuable in stable,
favorable environments with little change. Figure 2: Reproductive cycle of
Aspidoscelis. (Source: Campbell)

3 Fertilization
• In external fertilization, the female releases eggs into its surroundings, then the male
releases its sperm into the environment to fertilize the eggs. This form of fertilization requires
a moist environment.

– In spawning, for example, a population of individuals in the same area release their
gametes at the same time. Spawning is synchronized via environment cues, such as day
length, temperature, or pheromones.
– The palolo worm coordinates the release of gametes with the phase of the moon.
During the last quarter of the moon in the spring, palolo worms break off their gamete-
filled tails, which rise to the surface and release the egg. The eggs are then quickly
fertilized by sperm.
– Asynchronous fertilization is also possible. However, courtship behaviors encourage
the release of sperm and egg at the same time between two animals, leading to higher
fertilization success.

• Internal fertilization is more common in dry areas and requires complementary reproduc-
tive systems.

– Internal fertilization systems also make use of pheromones, small molecules similar to
hormones that alter the physiology of other organisms.

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– Animals that rely on internal fertilization often have fewer offspring, but those offspring
generally have higher survival rates. This is because zygotes and embryos are better
protected from potential predators, as they are kept safe from the external environment.

• Most animals have gonads in order to produce sperm and eggs for reproduction.

• Those lacking gonads, like palolo worms and other annelids, instead produce gametes from
undifferentiated cells lining the coelom.

• Many species also have accessory ducts and other

structures that nourish the sperm and optimize fer-
tilization.

– Fruit flies and other insects have complex repro-

ductive systems that include spermathecae,
which store sperm in females for months until re-
lease during optimal conditions. It is also worth
noting that female fruit flies expel old sperm and
replace it with new sperm when a second male
mates with them. As such, males may transfer
chemicals to the female after mating with her,
making her less receptive to mating again.
– In non-mammalian vertebrates, the digestive,
excretory, and reproductive systems all exit to
one opening, the cloaca. In these species, males
often turn their cloaca inside out to release their
sperm.
– In mammalian vertebrates, males often share
openings between the digestive and excretory
Figure 3: Sex organs in flies.
systems, while females often share openings be-
(Source: Campbell Biology)
tween the excretory and reproductive systems.

4 Development
4.1 General Development
• Although the Y chromosome is much smaller than the X chromosome, it is vital to male
development. The genes found on the Y chromosome are located in palindromic sequences,
allowing for local recombination, which protects the Y chromosome against mutations.

• The SRY gene on the Y chromosome codes for tissue-determining factor (TDF), which
begins the development of the male reproductive system around 40 days of development.
The lack of SRY results in the development of the ovaries instead.

• Within the testes are the seminiferous tubules, which appear early in the development of
the male reproductive system and consist of both non-germ and germ cells.

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– Leydig cells are the endocrine portion of the testes. In the presence of luteinizing
hormone (LH), they produce testosterone.
– Sertoli cells provide structural support and aid the development of sperm. They
help produce testosterone in the growing embryo, resulting in the masculinization of
embryonic tissues and contributing to the varying levels of testosterone throughout
growth.
∗ Testosterone increases 8 weeks after conception, falls in the second trimester, rises in
the third month after birth and falls in the 7-12 months after birth until adolescence.
– Germ cells become sperm (eggs in females) through meiosis and specialization.

4.2 Accessory Sex Organs and External Genitalia Development

• Male accessory organs develop from the Wolffian duct, while female accessory organs de-
velop from the Müllerian duct. Both of these ducts are present at birth.

– In males, Sertoli cells secrete Müllerian Inhibiting Factor (MIF), causing the
Müllerian duct to degenerate. Leydig cells also secrete testosterone, causing the Wolffian
duct to form the epididymis, vas deferens, seminal vesicles, and ejaculatory duct.
– In females, the lack of MIF results in degeneration of the Wolffian duct and the resulting
differentiation of the Müllerian duct into the accessory sex organs.

• In the first six weeks of development, males and females share the same external genitalia.
Testosterone is converted to a derivative called dihydrotestosterone (DHT), which mas-
culinizes the tissue and results in the external genitalia.

Figure 4: Pathway for development of sex organs. (Source: Fox Human Physiology)

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5 Endocrine Regulation
Puberty, gamete production, and maintenance of the sexual structures depends on the secretions
of the gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH).

• The production and secretion of gonadotropins depends on gonadotropin-releasing hor-

mone (GnRH).

• The ovaries and testies secrete inhibin, which inhibits FSH secretion. Furthermore, FSH and
LH induce sex steroid secretion, which in turn inhibits gonadotropin and GnRH secretion.
This is known as negative feedback.

• Gonadotropin secretion is pulsatile, thus preventing the desensitization of receptors.

• Kisspeptin, a neurotransmitter found in the hypothalamus, increases GnRH secretion. In

females, it changes the brain’s structure to induce cyclical GnRH secretion. In males, andro-
gens suppress kisspeptin secretion, allowing for noncyclical secretion.

• In many animals, puberty begins with the weakening of inhibitory receptors on the hypotha-
lamus. In humans, it is likely a result of decreased inhibition by GABAergic neurons, along
with increased kisspeptin secretion.

• In females, increased estradiol secretion results in the stimulation of the epiphyseal growth
plate and, thus, a growth spurt. Estrogen stimulates chondrocytes to divide, causing the
cartilage matrix to grow and calcify. Estrogen also stimulates breast development and causes
menarche, the first menstrual flow.

– Puberty is inhibited by exercise in females because leptin, secreted by adipose tissue, is

required for puberty.

• In males, testosterone results in the growth of the penis, as well as the development of
other sex characteristics. In bone, testosterone is converted to estrogen, stimulating growth.
Testosterone also directly stimulates bone growth under the periosteum, the membrane cov-
ering the bone. This results in wider bones.

• Hair growth in both males and females is promoted by steroids secreted by the adrenal cortex.

6 Male Reproductive System

6.1 Anatomy
• The scrotum and the penis constitute the external genitalia of the male.

• The internal male reproductive system consists of cells that secrete hormones and sperm. It
provides ducts for the movement of gametes and houses glands important for the production
of fluids necessary for the movement of sperm.

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– The testis consists of highly coiled seminiferous tubules involved in the production
of sperm. It must be kept at a temperature around two centigrade lower than the
core body temperature. This is attained through the lowering of the testis and the
surrounding scrotum during embryonic development such that they hang away from
the body. Cremaster muscles also travel through the spermatic cord to reflexively
move the testis up and down in order to maintain temperature. In rodents, the testis
ascends after mating season to prevent sperm production.

– Sperm moves from the seminiferous

tubules to the epididymis, where it can
stay for a few weeks. During ejac-
ulation, the sperm moves through a
muscular vas deferens, which connects
secretions from the seminal vesicle to
the ejaculatory duct. The ejaculatory
duct opens into the urethra, which
opens the tip of the penis.

• Accessory Glands

– The seminal vesicle, bulbourethral

gland, and prostate gland all secrete
fluids that combine with sperm to form
semen. Before ejaculation, the bul-
bourethral glands release a clear mu-
cus that neutralizes any acidic urine in
the way. The accessory glands’ func-
tions will be further described later in
this handout. Figure 5: Diagram of sperm’s pathway.
(Source: Campbell Biology)

Figure 6: Diagram of where sperm are formed. (Source: Fox Human Physiology)

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6.2 Gonadotropin Secretion

• Sertoli cells produce inhibin to inhibit FSH secretion. The latter is done when FSH itself
binds to receptors on Sertoli cells. This binding also stimulates spermatogenesis.

• Leydig cells produce testosterone to inhibit gonadotropin secretion. They are found in the
interstitium, which is heavily inundated with blood vessels in order to allow testosterone to
easily travel throughout the testis.

• Testosterone is converted to many different compounds within cells to function in different

tasks. Testosterone is converted to DHT in the prostate and skin and to estrogen in the
skeletal system. In males, estrogen is important in adipose tissue, while testosterone is
important in muscles.

View the table below to see the various actions of androgens (e.g., testosterone).

Figure 7: Synthesis pathway (left) and actions of (right) androgens.

(Source: Fox Human Physiology)

6.3 Spermatogenesis
• During embryonic development, diploid cells migrate to the embryonic testes and become
spermatagonia, stem cells that will produce sperm by mitosis and meiosis. These cells are
located by the basement membrane, closest to the nourishing blood vessels.

• The diploid cells undergo mitosis to form a primary spermatocyte while also regenerating
the original stem cell. This prevents the stem cell supply from being exhausted.

• The primary spermatocyte then undergoes meiosis to form secondary spermatocytes.

• The secondary spermatocytes undergo a second meiotic division to form spermatids.

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Figure 8: Spermiogenesis. (Source: Fox Human Physiology)

• The development of the spermatids into functional spermatozoa (i.e., sperm cells) requires a
process called spermiogenesis.

– During spermiogenesis, histones are modified and replaced by proteins called pro-
tamines. This allows for the compaction of the spermatozoa nucleus, followed by
the development of the flagella.
– Sertoli cells are located on the basement membrane and form a ring around the tubules
connected by tight junctions, a blood-testis barrier. This barrier prevents autoim-
mune reactions, only allowing certain molecules to pass. In other words, it creates an
immunologically privileged site, with little to no immune cells.
– Sertoli cells consist of large amounts of cytoplasm from the basement to the lumen.
They are connected via tight junctions between them, and spermatogenesis occurs in
the spaces between the tight junctions As the products slowly move through these
spaces, the tight junctions constantly break and reform to accommodate this change.
– During spermiogenesis, the Sertoli cells phagocytize the bulges of cytoplasm off the side
of the sperm. During this process, the Sertoli cells also provide chemicals and products
required by the sperm, such as products of the X chromosome not found in some sperm.
– Sertoli cells secrete androgen-binding protein (ABP) into the lumen of the tubules, al-
lowing for the concentration of testosterone within the tubule, where it aids the function
of sperm.

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• A spermatozoa consists of a head, with a nucleus, DNA, and an overlying cap called an
acrosome, as well as a three-part 9+2 flagella called an axoneme. The midpiece (superior)
contains mitochondria and a fibrous sheath. The principal piece only contains the fibrous
sheath. The endpiece (inferior) lacks both of these.

Figure 9: Spermatogenesis. (Source: Fox Human Physiology)

• In spermatogenesis, the division of primary spermatocytes past prophase I requires testos-

terone, which is converted to its derivatives and stimulates the completion of division. Testos-
terone secreted by Leydig cells also stimulates spermatogenesis in the seminiferous tubules.
• FSH is not required for spermatogenesis, but it makes for a more optimal environment and
allows for greater fertility.

6.4 Accessory Sex Organs

• Following sperm production in the seminiferous tubules, sperm is moved into the rete testis,
where it drains via the efferent ductules into the epididymis, which is then drained by the
vas deferens.
• When sperm enter the epididymis they are immotile due to the low pH. However, as they
move through the epididymis, they obtain resistance to this low pH.
– In the female reproductive tract, the pH is neutralized by the alkaline secretions of the
prostate, so sperm becomes fully active and motile.

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• The seminal vesicles are the first to add their fructose-rich fluid that composes 60% of the
volume in semen.

• Next, pores in the prostate gland allow citric acid, calcium, and coagulation proteins to enter
the semen.

– Coagulation proteins allow the semen to coagulate after ejection before being decoagu-
lated in the female reproductive system.

6.5 Erection, Emission, and Ejaculation

• The two corpora cavernosa consist of erectile tissue in the ventral portion of the penis.
The corpora cavernosa are innervated by the deep artery of the penis.

• Nitric oxide (NO), produced by parasympathetic axons, is released to the corpora cavernosa,
activating guanylyl cyclase (GC). This increases cyclic guanosine monophosphate (cGMP)
levels and causes Ca2+ outflow. This then results in vasodilation, which causes an erection.
Endothelial cells of the corpora cavernosa also produce NO, allowing for further vasodilation.

• The corpus spongiosum is a mass of erectile tissue surrounding the urethra in the dorsal
portion of the penis.

• Emission is the movement of semen into the urethra, and ejaculation is the expulsion of
semen from the urethra. Emission and ejaculation are stimulated by sympathetic nerves that
cause the contraction of muscles and glands.

7 Female Reproductive System

7.1 Anatomy
• The two ovaries, analogous to testes in males, enter into the Fallopian tube.

• The ova released into the uterine tubes by ovulation are drawn in by cilia into the uterus,
a muscular, pear-shaped organ. There are three layers in the uterus, the outer perimetrium,
middle myometrium, and the inner endometrium.

– The endometrium consists of nonkeratinized, stratified squamous epithelium with a

stratum basale and an upper stratum functionalis. The stratum functionalis grows
cyclically throughout the menstrual cycle.

• The uterus opens into the vagina, which is separated from the uterus by a cervical mucus.

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Figure 10: Diagram of female reproductive system. (Source: Fox Human Physiology)

7.2 Ovarian Cycle

• Germ cells migrate from the embryo and multiply via mitosis to form oogonia, most of which
undergo apoptosis. The remaining oogonia undergo meiosis and are arrested at prophase I
to form diploid primary oocytes, just as in spermatogenesis.

• Throughout the lifetime of a woman, only 400 oocytes will likely ovulate. This contrasts
with spermatogenesis, in which stem cells are regenerated. Primary oocytes that are not
stimulated to divide are contained in primary follicles.

• Primary follicles contain a single layer of surrounding follicle cells. FSH stimulation results
in division to form layers of granulosa cells that surround the oocyte.

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• The oocyte develops vesicles containing fluid, becoming a secondary follicle. The vesicles
eventually fuse and create a cavity called the antrum. This follicle is known as a Graafian
follicle.

• To form a secondary oocyte, the primary oocyte completes its meiotic division, forming a
polar body, which later degenerates, and a secondary oocyte, which retains most of the
cytoplasm. The secondary oocyte continues meiosis until it stops at metaphase II.

• The theca interna of the ovarian follicles produces testosterone in response to LH, which
diffuses into granulosa cells, where aromatase converts it into estradiol. Thus, estradiol is
produced as the follicles grow bigger.

Figure 11: Labeled cross-section of follicles. (Source: Fox Human Physiology)

• Usually, only one follicle becomes dominant and ovulates, while the rest are atrietic and
fail to rupture. FSH and LH promote ovulation and protect the follicles from atresia, while
androgens and certain proteins promote atresia.

• The mature follicle ruptures via ovulation, still surrounded by the corona radiata and zona
pellucida. The sperm must pass through these layers to fertilize the egg and complete the
last meiotic division, which produces a polar body and developing zygote.

• Following ovulation, LH promotes the formation of the corpus lutetum out of the empty
follicle, which secretes estradiol and progesterone. At the end of menstruation, it regresses
to form a non-functional corpus albicans.

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Figure 12: Diagram of follicle development. (Source: Fox Human Physiology)

7.3 Menstrual Cycle

• In females, cyclic changes in gonadotropic hormones result in ovarian changes that follow
monthly cycles. The resulting hormonal changes are accompanied by changes in the en-
dometrium.
• Only primates have a menstrual cycle. In other mammals, the endometrium is simply reab-
sorbed without fluid flow. Mammals with estrous cycles only undergo sexual activity during
the period surrounding ovulation in order to maximize pregnancy chances, while those with
menstrual cycles can copulate throughout the year. Bears and wolves have a single estrous
cycle, while elephants have several. Cats ovulate only upon mating.
• The follicular phase lasts from the first day of menstruation until ovulation (i.e., days
1-13). Menstruation lasts from days 1-5 of an average cycle.
– As follicles grow, becoming Graafian follicles, estradiol levels begin to rise, reaching
their peak at about day 13. Low levels of estradiol inhibit LH, However, high levels of
estradiol stimulate the secretion of LH.
– FSH stimulates the production of FSH receptors on granulosa cells, increasing their
sensitivity. FSH and estradiol also stimulate the production of LH receptors.
– FSH levels decline around the middle of the menstrual cycle, causing atresia of non-
dominant follicles. However, the increased sensitivity of FSH receptors and the increased
number of FSH receptors allow dominant follicles to survive this fall in FSH.
– The rise in estradiol during the follicular phase increases GnRH pulses via positive
feedback. As a result, LH levels increase rapidly, generating an LH surge that triggers
ovulation.

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• The luteal phase lasts from the first day of ovulation until the first day of menstruation.
– After ovulation, LH stimulates the empty follicle to develop into the corpus luteum.
This structure secretes progesterone and estradiol, allowing for negative feedback on
FSH and LH secretion. This prevents the development of new follicles in order to avoid
multiple simultaneous pregnancies. Note that the collective action of estradiol and
progesterone results in negative feedback, while estradiol by itself results in positive
feedback.
– The late luteal phase sees a drop in estrogen and progesterone levels as the corpus
luteum becomes the corpus albicans. This transition is mediated by a uterine hormone
called luteolysin. Normal function of this hormone is prevented by LH, but the corpus
luteum lowers LH levels, leading to its own degradation.
– At the end of the cycle, estrogen and progesterone levels fall, allowing a new cycle to
begin.

Figure 13: Hormone levels throughout the menstrual cycle. (Source: Fox Human Physiology)

7.4 Changes in the Endometrium

• The proliferative phase coincides with the follicular phase of menstruation. Increased
estradiol levels stimulate the growth of the stratum functionalis, along with coiled blood
vessels in the endometrium, found only in primates, called spiral arteries.
• The secretory phase coincides with the luteal phase of menstruation. Increased proges-
terone levels stimulate the development of uterine glands, resulting in a thick and spongy
endometrium. This phase also sees an increase in glycogen levels in the endometrium.
• The menstrual phase sees necrosis of the stratum functionalis. Furthermore, vasoconstric-
tion of arteries results in bleeding and shedding of the endometrium.
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7.5 Menopause
• The depletion of follicles results in a decrease in estrogen and inhibin secretion, which in-
creases FSH and LH secretion due to the loss of negative feedback. The lack of estrogen
often leads to osteoporosis in postmenopausal women.

• Postmenopausal women have small amounts of estrone, a type of estrogen that is formed by
androgens in adipose tissue.

• Women with higher amounts of adipose later in life, therefore, have a lower chance of devel-
oping osteoporosis.

• Menopause causes many symptoms, including loss of lubrication in the vaginal wall associated
with atrophy, increased risk of atherosclerosis, osteoporosis, and hot flashes (i.e., falls in core
body temperature, followed by sudden feelings of heat and sweating).

Example 7.1: (USABO Semifinals Exam 2020) With the onset of menopause, ovarian
follicles gradually stop releasing eggs. A corresponding change in reproductive hormone
levels occurs. The figure shows the changes in estradiol, FSH, hormone X, and LH during
this pre- to post-menopausal transition. Hormone X is a marker for developing follicles.
Which choice correctly labels the four hormones?

a. I - Estradiol; II - Hormone X; III - FSH; IV - LH

b. I - FSH; II - LH; III - Estradiol; IV - Hormone X
c. I - FSH; II - Estradiol; III - LH; IV - Hormone X
d. I - Hormone X; II - LH; III - Estradiol; IV - FSH
e. I - Estradiol; II - Hormone X; III - LH; IV - FSH

Solution: During menopause, follicles become depleted, and estrogen and inhibin secretion also
fall. This results in increased FSH and LH secretion, meaning they correspond to I and II on the
graph. Since follicles are depleted, hormone X must be either III or IV, leaving estradiol to also
be III or IV. Thus, the answer must be B.

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8 Fertilization, Lactation, and Birth

8.1 Capacitation
• The male ejaculates around 300 million sperm into the female vagina. Only around 100
survive, of which about 10 gain the ability to fertilize an ovum, known as capacitance.

• The female reproductive tract is alkaline. In conjunction with the removal of hydrogen from
the sperm, this allows for a raise in the pH of a sperm cytoplasm and thus the activation of
dynein, a motor protein, in the flagellum.

• The rise in sperm pH also activates a calcium channel in the flagellum’s principal piece,
called a CatSper channel. The subsequent rise in calcium results in hyperactivation of the
flagellum, with a resulting increase in motility.

• The sperm moves along the oviduct via chemotaxis and thermotaxis, attracted toward specific
chemicals and warmer temperatures.

8.2 Fertilization
• Sperm binds to carbohydrates on the glycoprotein-rich zona pellucida. Through this binding,
sperm is exposed to progesterone, which is secreted by the corona radiata around the ovum.

• The progesterone activates CatSper channels in the head of the sperm, inducing the acro-
some reaction, in which the acrosome in the sperm fuses with the outer membrane, resulting
in the release of acrosomal enzymes. These enzymes, which are proteases and hyaluronidases,
digest the hyaluronic acid surrounding the extracellular matrix of the egg, allowing the sperm
to dig through the zona pellucida to fuse with the egg.

• When the sperm binds to the egg, a calcium wave is triggered, in which cytoplasmic calcium
increases, starting from one pole of the egg and traveling to the opposite pole. This wave
prevents other sperm from fertilizing the egg and also restarts the second meiotic division.

• The sperm contributes DNA and a centrosome, necessary for microtubule organization during
mitosis. Mitochondria brought in by the sperm degenerate. Thus, all mitochondrial DNA is
maternal.

• In in vitro fertilization (IVF), the mother receives injections of FSH to stimulate the
growth of ovarian follicles, along with other hormones to prevent ovulation. The follicles are
removed from the ovary, and sperm is injected into the egg through the zona pellucida. The
embryo is then grown until it reaches the 8-cell stage, in which it is transferred to the uterus
to restart growth.

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Figure 14: Diagram of fertilization. (Source: Fox Human Physiology)

8.3 Birth and Pregnancy

• After ovulation, a sperm fertilizes the oocyte and restarts meiosis. Cleavage occurs as the
egg travels through the oviduct. By the time the embryo reaches the uterus, it is a ball of
cells. The embryo then implants itself into the uterus about seven days after conception.

• In a typical menstrual cycle, the corpus luteum would degenerate, causing a drop in hormones
that makes the endometrium slough off. In order to prevent this in a pregnant female, the
implanted embryo secretes human chorionic gonadotropin (hCG), similar to LH, in order
to maintain the corpus luteum so that it continues to secrete progesterone and estrogen.

• Carrying an embryo in the stomach is known as pregnancy or gestation.

• During the first trimester, which lasts for three months, the embryo secretes hCG to
maintain the secretion of progesterone and estrogen by the corpus luteum. Like other go-
nadotropins, hCG can be detected in or extracted from the urine.

– The embryo obtains its nutrients from the endometrium, while the outer trophoblast
forms outgrowths and associates with the endometrium to form the placenta.
– The placenta provides nutrients and immune protection to the embryo and helps it
dispose of metabolic wastes. The umbilical cord connects the placenta with the fetus,
allowing blood to travel between the two.
– If an embryo splits during the first trimester, it can form a monozygotic (i.e., identical)
twin. If two follicles mature and are both fertilized and implanted, then dizygotic (i.e.,
fraternal) twins can be born.
– During organogenesis, the development of body organs, the embryo is very susceptible
to alcohol and other damage. The heart begins to beat by the 4th week and can be
detected by the 8th-10th week using a handheld Doppler.

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– Hormonal changes such as high levels of progesterone result in changes in the mother,
including the formation of a mucus plug in the cervix, enlargement of the breasts and
uterus, and interruption of menstrual cycling. The mother may also experience nausea
and morning sickness.
– By the end of the first trimester, the embryo is about 5 centimeters long.

• The second and third trimesters see the formation of fingernails, external genitalia, and
the outer ears, as well as the beginning of fetal movements.

– The corpus luteum degenerates, and the placenta takes over the production of go-
nadotropins.
– By the end of the second trimester, the embryo grows to about 30 centimeters long.
– In the third trimester, the fetus grows to about 50 centimeters long, and activity slows
down as the fetus fills the amniotic sac.
– Labor follows 3 stages: dilation of the cervix, delivery of the baby, and delivery of
the placenta. During labor, contractions are induced in the uterus by local regulators
and hormones to push the fetus and placenta out of the body. Uterine contractions
stimulate oxytocin secretion, providing positive feedback to continue contractions.
– In the fetus, ACTH is released by the posterior pituitary in response to CRH from
the hypothalamus, resulting in the secretion of cortisol and dehydroepiandrosterone
(DHEA), an androgen, from the adrenal cortex.
– Cortisol results in surfactant secretion to prepare the fetal lungs to breathe.
– DHEA is converted to estriol, which activates the myometrium to express more receptors
for oxytocin and prostaglandins. Typically, progesterone inhibits this function of estriol.
In addition, estriol stimulates gap junction formation between myometrial cells to allow
for synchronous contraction.

Figure 15: Positive feedback loop during birth. (Source: Campbell Biology)

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Figure 16: Detailed pathway of the positive feedback in birth. (Source: Fox Human Physiology)

8.4 Lactation
• Mammary glands are composed of multiple lobules, within which lies around 15 lobes in
each breast. The lobules contain structures known as alveoli, different from those of the
lungs, that secrete milk into ducts that lead up to the nipple.

• Prolactin stimulates the production of milk by the mammary glands. Dopamine inhibits
prolactin production, and estrogen stimulates dopamine production.

• When the placenta is expelled after labor, the rapid drop in estrogen results in an increase
in prolactin that stimulates milk production.

• Breastfeeding results in the nursing reflex, which inhibits dopamine secretion and results in
oxytocin secretion. Oxytocin stimulates milk ejection via myoepithelial cells, which are
involved in the propulsion of milk through the ducts to the nipple. This reflex also inhibits
GnRH secretion and is thus a natural contraceptive.

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9 Disorders
9.1 Hermaphroditism and Pseudohermaphroditism
• Hermaphroditism occurs when some embryonic cells obtains the short arm of the Y chro-
mosome while others do note, this sometimes results in half ovary half testis or ovotestis
(ovary and testis combined).

• Pseudohermaphroditism is characterized by underdeveloped or incorrect external geni-

talia that does not match with their gender.

– Congenital Adrenal Hyperplasia occurs when there is a mutation in cortisol syn-

thesis resulting in loss of negative feedback (due to reduced cortisol) and thus large
amounts of androgen secretion from the adrenal cortex. Females will still develop a
uterus but the external genitalia will be ambiguous and masculinized.
– Testicular Feminization Syndrome occurs when testosterone receptors are faulty
leading to feminized genitalia. Furthermore, there is a lack of negative feedback leading
to large amounts of circulating testosterone. This testosterone gets converted to estrogen
in adipose tissues which results in a female appearance without menstruation.
– 5-alpha Reductase Deficiency occurs when the enzyme 5--reductase is deficient
which leads to normal internal genitalia but underdeveloped external genitalia. This is
because 5--reductase is used to make DHT from testosterone, and DHT is required for
external genitalia formation.

9.2 Endometriosis
• Endometriosis results from the back flow of endometrial tissue through the uterine tubes into
the pelvic cavity where it responds to cycles in hormones and undergoes normal menstruation
and bleeding. This results in scarring, inflammation, infertility and intense pain.

• Treated with a drug called nafarelin that acts as a GnRH analogue resulting in continuous
hormonal secretion which as opposed to normal pulsatile secretion results in downregulation
of GnRH receptors and thus decreased LH and FSH secretion. This allows for menopause
like conditions that prevents menstruation and thus abnormal bleeding.

9.3 Ectopic Pregnancy

• An ectopic pregnancy occurs when a fertilized egg implants within the Fallopian tube result-
ing in internal bleeding via Fallopian tube rupture.

– The chance of this condition occurring becomes more likely due to bacterial scarring
from, for example, an STI (sexually transmitted infection).

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9.4 Genetic Screening

• Chorionic Villus Sampling, a needle is inserted through the cervix and a portion of the
placenta (composed of the fetus’ cells) is removed. These cells proliferate fast, so karyotype
and molecular tests can occur immediately. Can be done after the 10th week of pregnancy.

• Amniocentesis, a needle is inserted through the uterus and a sample of amniotic fluid
is taken and tested, an ultrasound monitors this process. This fluid must be centrifuged to
isolate the cells and then can be tested for certain molecules indicative of a disease, otherwise
they must be cultured for several weeks before preforming a karyotype on them. Only can
be done after the 15th week of pregnancy.

• Ultrasound

– Reflected sound waves are used to project an image of the fetus, noninvasive and allows
for the imaging of anatomical abnormalities.

Figure 17: (Source: Campbell Biology)

10 Contraceptives
Contraception is the prevention of pregnancy. There are many contraceptives and many opportu-
nities for the prevention of pregnancy. The most effective contraceptives are intrauterine devices
(IUDs) and hormonal contraceptives.

10.1 Rhythm Method

• Conception is most likely to occur one to two days before ovulation. Therefore natural
family planning, the rhythm method, or temporary abstinence involves planned
sexual intercourse times to lower the likelihood of pregnancy. More than 6 days before and
one day after ovulation are the safest times to have coitus.

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• Temperature changes during the menstrual cycle can be tracked. There is a lowered tem-
perature during the LH surge due to low estradiol levels. However, temperature increases
after ovulation as progesterone increases. Monitoring body temperature is thus useful for a
woman who is planning to have kids but not to prevent pregnancy, as it cannot be used to
predict menstruation.

10.2 Coitus Interrupts

• This method involves the withdrawal of the penis prior to ejaculation. This method is highly
unreliable, as the bulbourethral gland, also known as Cowper’s gland, often releases sperm
when clearing the urethra of urine.

10.3 Oral Contraceptives

• Oral contraceptives increase estradiol and pro-
gesterone levels using a synthetic estrogen and a
synthetic progesterone known as progestin. This
results in negative feedback of GnRH and thus
lowered FSH and LH secretion, preventing ovu-
lation. Progestin by itself results in the thicken-
ing of cervical mucus to block sperm from enter-
ing the uterus.

• To prevent abnormal endometrium growth,

these pills are stopped on the third week to allow
for menstruation, then restarted with a placebo
pill for the fourth week before continuing.

• Patches and oral contraceptives prevent drug

metabolism by the liver and include the trans-
dermal patch, vaginal ring, and injections of
medroxyprogesterone acetate (DMPA). IUDs
and subcutaneous patches are more long-acting
than the contraceptives listed above. Further-
more, these contraceptives may increase the
chance of heart attack.

10.4 Sterilization
• Sterilization is the prevention of gamete release.
Tubal ligation, one method of this, is the ty-
ing of the oviduct to prevent eggs from travel-
ing. Vasectomy, another method, is the cut-
ting and knotting of the vas deferens to prevent
Figure 18: Points of contraception.
sperm from entering during ejaculation. In a va-
(Source: Campbell Biology)
sectomized male, sperm does not build up; it
is reabsorbed when it reaches the knot. These
methods do not affect sex hormone secretion.

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10.5 Abortion
• Abortion is the termination of a pregnancy in progress. Mifepristone (RU486) blocks
progesterone receptors in the uterus, preventing the maintenance of pregnancy. It is often
taken with prostaglandins to induce uterine contraction.

11 Development
11.1 Cleavage
• Cleavage is the rapid cell division that occurs in a newly fertilized egg. It consists of mainly
the S and M phases without growth during the G1 and G2 phases.
• Development of a fertilized egg is carried out by preexisting mRNA and proteins, as the
DNA in the egg is insufficient to sustain such a large cytoplasm.
• There is no increase in the size of the cells. Rather, the cytoplasm is partitioned into
blastomeres, each of which consists of a blastula surrounding a fluid-filled cavity called a
blastocoel.
• In species such as sea urchins, cleavage is uniform across embryos. In frogs and other species,
it is asymmetric.

11.2 Gastrulation in Frogs

• Each germ layer corresponds to a different set of structures in the adult animal. For example,
the ectoderm forms the outer nervous system, while the mesoderm gives rise to the muscle
and skeleton.

Figure 19: The products of the three germ layers. (Source: [Link])

• In frogs, gastrulation begins on the dorsal side, opposite to the entry of sperm. Gastrulation
generates three germ layers. In other words, frogs, like vertebrates and other bilaterally
symmetric animals, are triploblasts. Diploblasts only have two germ layers. The extra
germ layer in a triploblast is known as the mesoderm.
• First, cells on the dorsal side invaginate and form a crease known as the blastopore.
Cells from the animal pole begin to roll into the hollow interior, forming the mesoderm and
endoderm of the gastrula.

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• Next, the blastospore spreads along the cell, and the two ends connect to form a circle that
becomes smaller as the ectoderm moves downward.

• finally, the germ layers have been created and now begin development. The blastospore has
become a yolk plug

Figure 20: Diagram of steps in gastrulation. (Source: Campbell Biology)

11.3 Gastrulation in Chicks

• There are many similarities between chicken and frog gastrulation. As you read on, try to
make connections between them as a result of the conserved evolution of development.

• A chicken egg is composed of the yellow that we commonly call yolk. A small spot on top of
this yolk is the animal pole, which will actually form the embryo.

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• The cells of the epiblast are the ones that eventually form the embryo of the chicken. These
cells move towards the midline of the animal pole in a process analogous to the dorsal lip in
amphibians, forming a furrow called the primitive streak.

• The cells that form the primitive streak push the lower hypoblast cells to form the endoderm,
while the rest form the mesoderm. The cells left at the top form the ectoderm.

• The cells of the hypoblast connect the yolk mass to the embryo and form a sac around the
yolk.

Example 11.1: (USABO Open Exam 2017) Select all of the following choices that correctly
match the tissue to the embryonic germ layer from which it is primarily formed (Select ALL
that apply):

A. Ectoderm and spinal cord.

B. Mesoderm and heart.
C. Endoderm and thyroid.
D. Ectoderm and epidermis.
E. Endoderm and liver.

Solution: The spinal cord forms from the neural tube, which is derived from ectodermal tissue.
The heart is part of the circulatory system, which is derived from the mesoderm. Remember
that in the previous diagram, the thyroid was described to be derived from the endoderm. The
epidermis is the top layer of the skin, while the dermis is below it. Thus, the epidermis is derived
from the ectoderm, while the dermis is derived from the mesoderm. Finally, the liver is part of the
digestive system, which is derived from the endodermal germ layer. The answer is ABCDE.

11.4 Gastrulation in Humans

• In humans, fertilization occurs in the oviduct, and development begins as the egg makes its
way down the oviduct.

• Before implantation, the embryo has an inner cell mass that eventually develops into the
fetus, as well as a surrounding chorion composed of trophoblast cells.

• The chorion develops into an outer syncytiotrophoblast and an inner cytotrophoblast. The
inner cell mass that gives rise to the 3 germ layers is separated from the chorion by the
amniotic cavity.

• The syncytiotrophoblast secretes enzymes that digest the endometrium, creating blood-filled
cavities into which the cytotrophoblast extend villi. This forms the chorion frondosum,
which eventually forms the placenta. The placenta consists of the fetal frondosum and the
maternal decidua basalis in contact with the frondosum.

• The trophoblast forms the 4 extraembryonic membranes that enclose structures outside of
the embryo. Gastrulation begins and a primitive streak forms, just as in chicks, with some
epiblast cells forming the mesoderm and endoderm and other cells forming the ectoderm.

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• The extraembryonic mesoderm and the 4 extraembryonic membranes surround the em-
bryo. In mammals, reptiles, and birds, these 4 extraembryonic membranes are the chorion,
allantois, amnion, and yolk sac. All vertebrate embryos require an aqueous environment to
allow for development.

– In amniotes, which include mammals, reptiles, and birds, a container shell or uterus
allows for developing embryos to be surrounded by a fluid called the amnion. Fish and
amphibians do not require an amnion because their eggs are surrounded by water.
– The chorion functions in gas exchange. The amnion protects the embryo. The allantois
disposes of waste in reptile eggs, and it does the same as part of the umbilical cord in
mammals. The yolk sac encloses the yolk in reptile eggs.

Figure 21: Implanting of the embryo into the placenta. (Source: Fox Human Physiology)

11.5 Organogenesis
• During organogenesis, the embryonic germ layers develop into the organs. Sometimes, mul-
tiple germ layers form different parts of a single organ.

• In neurulation, cells from the mesoderm form the notochord, which runs down the dorsal
side of the embryo. These cells secrete signaling molecules that cause the ectoderm above to
form the neural plate. The cells of the neural plate curve the structure inward to form a
neural tube. The anterior end develops into the brain, while the posterior end forms the
spinal cord. The notochord degenerates except in certain parts that form the disks in the
spine.

– Spina bifida is the failure of the neural tube to close properly. It can result in nerve
damage and thus leg paralysis.

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• In cell migration, some cells develop on the edges where the neural tube pinches off from
the neural fold. These cells migrate to the periphery of the embryo forming nerves, teeth,
and skull bones. Cells lateral to the notochord form blocks known as somites, which organize
the segmentation of the body. Some form vertebrae, skeletal muscles, and mesenchyme, the
latter of which consists of stem cells in the skeletal system.

Figure 22: Organogenesis in chicks. (Source: Campbell Biology)

11.6 Cytoskeleton in Morphogenesis

• In neurulation, microtubules orient themselves laterally at the ventral surface of the cuboidal
ectoderm and horizontally at the dorsal surface. Actin contraction creates a wedge shape
that bends the ectoderm inward.

• In convergent extension, cells elongate in one direction and crawl between each other.
Thus, the sheet becomes long and narrow. This process occurs in the formation of the
primitive streak of chick eggs and the archenteron of sea urchin embryos.

• In cell migration, cells of the neural crest and somites migrate throughout the embryo via
the cytoskeleton, similar to ameboid movement. Cell adhesion molecules (i.e., glycoproteins)
are important in this process, as well as the extracellular matrix (ECM).

11.7 Fate Mapping

• In fate mapping, specific regions of a blastula are marked with a dye, and the location of the
cells in those regions is tracked to study the origin of adult tissues.

• Microscopic analysis can also be used to track cells, along with mutating or destroying cells
with a focused laser and observing its effect on development.

• Fate mapping was used to determine the location of P granules in C. elegans. They were
found to move posteriorly during the first mitotic cell division and become concentrated
toward the end of cleavage, eventually giving rise to the germline.

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11.8 Axis Formation

• In frogs, the anterior-posterior (AP) axis is formed during oogenesis. However, the dorsal-
ventral (DV) axis is formed after fertilization. When the sperm binds to the animal pole,
the overlying cytoplasm rotates toward the point of entry. This activates certain regulatory
proteins that allow the dorsal side to form in the gray crescent, an area formed by the
rotation that is opposite to where the sperm entered.

• In humans, the orientation of the sperm’s nucleus with respect to the egg’s nucleus determines
the orientation of cleavage. Both axes form in the blastula and gastrula stages.

• In chickens, the pull of gravity as the egg travels down the oviduct determines the AP axis.

• In zebrafish, signals across the embryo determine axial patterning.

• In insects, gradients of transcription factors determine axial patterning.

• Once the AP and DV axes have been determined, the left-right (LR) axis is easy to determine.
In the two-cilia hypothesis for vertebrates, motile cilia rotate clockwise to drive fluid to
flow to the left, while non-motile cilia detect this flow. This allows for left-right asymmetry.

11.9 Induction in Pattern Formation

• Pattern formation is the arrangement of organs in the place they should be. This is brought
upon by positional information, a cell’s location in relation to the defined axes.

• A scientist and his student (i.e., Spemann and Mangold) transplanted a dorsal lip from one
frog onto another part of a different frog, triggering gastrulation. Later, the Spemann
organizer was named, a group of cells that inhibit bone morphogenic protein 4 (BMP4),
which normally prevents neural tube and notochord formation.

• In chickens, wings and limbs form as limb buds, which is composed of a mesoderm tissue
covered by overlying ectoderm. The anterior side of the limb is toward the thumb, while the
posterior side is toward the little finger. The dorsal side is the back of the hand, while the
ventral side is the palm side. The proximal side is toward the shoulder, while the distal side
is toward the hand.

• The apical ectodermal ridge (AER) on the ectoderm of the limb bud secretes fibroblast
growth factor (FGF), which promotes limb outgrowth along the proximal-distal axis.

• The zone of polarizing activity (ZPA) is found on the posterior side of the mesoderm of
the limb bud. It secretes a protein called sonic hedgehog (SHH), whose gradient determines
the formation of the digits. At high concentrations nearest the ZPA, SHH induces the
formation of the posterior digits (e.g., the pinky). At low concentrations farthest from the
ZPA, SHH induces the formation of the anterior digits (e.g., the thumb).

Example 11.2 (USABO Semifinal Exam 2018) Which of the following correctly lists the
steps required for the formation of neural tubes in vertebrates?
I. Cuboidal ectodermal cells form a continuous sheet.

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II. Microtubules help elongate the cells of the neural plate.

III. Actin filaments at the dorsal end of the cells may contract, deforming the cells into
wedge cells.
IV. Cells wedging in the opposite direction causes the ectoderm to form a hinge. It pinches
off of the neural plate and forms the neural tube.

A. I → II → III → IV
B. I → III → II → IV
C. II → I → III → IV
D. I → II → IV → III
E. III → II → I → IV

Solution: Looking through the choices, we know that IV must be last, as it includes ”forms the
neural tube.” We now look at the rest of the answer choices. During neurulation, the ectoderm is
composed of cuboidal cells that overlie the epithelium. Thus, I has to be the first step. As discussed
in this handout, actin contraction leads to the wedging of the neural plate to form the neural tube.
Therefore, III must be the second to last step, and II must be the second step. Logically speaking,
this also makes sense, as the cell must be elongated and have more microtubules to contract. Thus,
the answer is A.

12 Conclusion
As we conclude this comprehensive journey through the realms of development and reproduction,
we know now more about life’s most intricate and enduring processes. From the inception of life
through the miracle of fertilization to the intricate dance of cellular divisions, and the orchestration
of growth, we have uncovered the underlying mechanisms that govern existence. I hope this
handout served you well and aids you on your USABO endeavors.
Peace Out - Zelmay Jan

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