Exercise 2

Gonads and Gametogenesis

Names: Abas M., Balabagan M., Biñan A., Blanco M., & Sy V. L. Date: 02/13/24
Subject and Section: Developmental Biology / I2813 Score:

I. Ovary:
Label all the parts (based on the list above) and answer the questions.

A. Cat Ovary

1. Cat ovary 5X (cortical primordial follicles, cortex, medulla)

2. Cat ovary (tunica albuginea, earliest primordial follicles, oocyte cytoplasm,

nucleus, zona pellucida, follicle cells, stromal cells)
3. Cat ovary (zona pellucida, antrum, secondary follicle, theca of follicle, cortical
stroma, nucleus, blood vessel)

4. Cat ovary 250x (germinal epithelium, cortical stroma, primordial follicles, blood
vessel)

5. Cat ovary (secondary follicle, theca of follicle, zona pellucida, nucleus, antrum,
cortical stroma blood vessel)
6. Cat ovary 125x (Germinal epithelium, cortical stroma, theca of follicle, blood
vessel, secondary follicle, nucleus, antrum)

7. Cat ovary (atretic follicle, theca interna cells, theca externa cells, corpora radiata,
zona pellucida, primary oocyte, follicular antrum, zona granulosa, primordial
follicle, basement membrane)

8. Cat ovary (theca, granulosa cells, cumulus oophorus, oocyte, nucleus,

cytoplasm, follicle antrum, zona pellucida)
9. Frog immature ovary 32x (germinal epithelium, ovarian follicles, ovarian cavity)

10. Frog ovary (ovarian cavity, follicular cells, nuclei, yolk granules, oocyte,
connective tissue, germinal epithelium)

11. Frog mature ovary 64X (vegetal pole, animal pole, cortex, oocyte)
II. Testis
Label all the parts and answer the questions.

A. Mouse/Rat/Human testis

1. Mouse testis (testis, visceral tunica vaginalis, epididymis)

testis visceral tunica vaginalis

ansci.wisc.edu
Epididymis

2. Mouse testis (epididymis, adipose tissue, visceral tunica vaginalis,

seminiferous tubules, Leydig cells)

ansci.wisc.edu
3. Rat testis (tunica albuginea, blood vessel, seminiferous tubules, lumen)

it.stlawu.edu/
4. Rat seminiferous tubules (basement membrane of the seminiferous tubules,
spermatogonia)

cvhs.okstate.edu
5. Rat seminiferous tubules (primary spermatocytes, spermatogonia)

cvhs.okstate.edu
6. Rat seminiferous tubules (early spermatids, late spermatids)

cvhs.okstate.edu

7. Rat seminiferous tubules (spermatids, Sertoli cells, Sertoli cell cytoplasm)

cvhs.okstate.edu
8. Human testis (primary spermatocyte, Sertoli cells, spermatids maturation
phase, spermatogonia, smooth muscle)

med.unsw.edu.au

9. Human testis (Leydig cells, Sertoli cells, spermatogonia, spermatids, primary

spermatocytes, smooth muscle)

med.unsw.edu.au
B. Frog testis

1. Xenopus laevis adult testis (interstitium, seminiferous tubules, tunica albuginea, vas
efferens)

Wiley

2. Xenopus laevis adult testis (lumen of the seminiferous tubule, primary

spermatogonium, spermatic duct, secondary spermatogonium, seminiferous tubule,
spermatozoa)

Wiley
3. Xenopus laevis adult testis (seminiferous tubule, discharging spermatozoa, spermatic
duct)

Wiley

III. Sperm
A. Rat sperm cell 1000X (head, flagella, midpiece)

Researchgate
 B. Human sperm cell (head, flagella, midpiece)

Britannica

Questions:

1. What is the functional significance of the numerous nucleoli of the

oocytes? Of the yolk?
Oocytes contain many nucleoli, which have multiple functional purposes.
First of all, they aid in the production of proteins necessary for the early stages
of embryonic development by participating in ribosome biogenesis. Ribosomal
RNA (rRNA) and ribosomal proteins, which are essential parts of
ribosomes—the cellular apparatus in charge of protein synthesis—are found in
nucleoli. Nucleoli also assist in the synthesis of ribosomal subunits, which
guarantees the availability of enough ribosomes to sustain fast cell division and
expansion throughout embryogenesis. Additionally, nucleoli support the general
development and viability of the embryo by controlling cellular processes as
aging, stress responses, and cell cycle progression (Boisvert et al., 2007;
Hernandez-Verdun, 2011).
The yolk found in oocytes acts as a store of nutrients for the growing
embryo, supplying proteins, lipids, and carbohydrates—essential
macromolecules needed for cellular growth, metabolism, and differentiation.
When a species develops lecithotrophically—that is, when the embryo depends
only on the mother's nutrients stored in the yolk to sustain itself until hatching or
birth—yolk becomes very significant. Antioxidants, vitamins, and other bioactive
compounds found in yolk components also support the health and viability of
the developing embryo. Depending on the species, the amount and distribution
of yolk in the egg can affect the growth and survival of the embryo (Wolfe &
Botelho, 2021).
2. What is the functional significance of lighter-staining cells (e.,
granulosa lutein cells) from those of darker-staining cells (theca lutein
cells)?
The ovary's darker-staining theca lutein cells and lighter-staining
granulosa lutein cells work in cooperation to control ovarian activity and
hormone synthesis. The ovarian follicle's granulosa lutein cells are principally in
charge of producing estrogen, a hormone vital to the growth and upkeep of the
female reproductive system. The growth and maturity of ovarian follicles, the
proliferation of the uterine endometrial lining, and the regulation of the
menstrual cycle are all significantly aided by estrogen (Richards, 2001). The
formation of the corpus luteum, which generates progesterone during the luteal
phase of the menstrual cycle and aids in endometrial preparation for
implantation and the maintenance of early pregnancy, is another function of
granulosa lutein cells (Richards, 2001; Speroff et al., 2010).
In contrast, androgens, which are precursors of estrogen biosynthesis,
are produced by theca lutein cells, which are found outside of the follicle. In
response to luteinizing hormone (LH), theca lutein cells synthesize androgens
like testosterone, which are then transformed into estrogen by granulosa cells
(Richards, 2001). Theca lutein cells generate androgens, which are essential for
follicular development and ovulation. They facilitate the maturation and
expansion of ovarian follicles and encourage granulosa cells to synthesize
estrogen (Speroff et al., 2010).
The preservation of female reproductive health and fertility is facilitated
by the coordinated action of granulosa lutein cells and theca lutein cells, which
guarantees the regulation of ovarian function, follicular development, and
hormone synthesis.

3. Differentiate a primary from a secondary follicle in terms of histological

appearance.
Histological appearance and developmental stage within the ovary can
be used to distinguish between primary and secondary follicles. A single layer of
cuboidal or flattened follicle cells (granulosa cells) encircling the oocyte is what
distinguishes primary follicles. Usually located in the ovary's cortex, these
follicles are in the early phases of development. A primary follicle's oocyte is
rather big and has a noticeable nucleus, or germminal vesicle, encircled by
cytoplasm. Compared to main follicles, secondary follicles are more developed
and have extra structural characteristics. A characteristic that sets secondary
follicles apart is the antrum, which is a cavity filled with fluid that encircles the
egg. The growth of granulosa cells and the creation of several layers (stratum
granulosum) around the oocyte are linked to the development of the antrum.
Furthermore, a more distinct basement membrane (zona pellucida) that isolates
the oocyte from the surrounding granulosa cells may be seen in secondary
follicles (Ceder & Collins, 2003).
4. Of what importance is the liquor folliculi to the oocyte?
The liquor folliculi, also known as follicular fluid, is filled with a liquid that
envelops the oocytes inside the ovarian follicle. It has a lot of significance for the
oocytes, which includes life support for maturing oocytes and feeding them with
amino acids, sugars, vitamins, and growth factors. a trash collector for maturing
oocytes, removing waste, and keeping things clean for healthy development.
Acts as a hormone highway, shuttling messages between granulosa cells
(making estrogen and progesterone) and the oocyte for smooth development
and growth control. Protection and support system for the oocyte, protecting it
from bumps and keeping the whole ovarian follicle nice and stable. lastly,
facilitates the process of fertilization.

5. Why are atretic follicles more numerous than the other cells? What are
the causes of atresia?
Atretic follicles, or follicles that undergo degeneration and regression, are
more numerous compared to other follicles due to a natural process of follicular
selection and elimination. There are several reasons why atretic follicles are
more numerous, for instance, competition and selection, whereas many follicles
begin to form throughout each menstrual cycle, yet a "chosen one" gets the best
food and hormones while the others fade away (atresia). It's a competition to
ensure the healthiest egg cell has the potential for fertilization.Limited
resources: the ovary cannot support all developing follicles simultaneously.
Follicles with lower responsiveness to essential hormones like FSH and LH
struggle to compete and ultimately degenerate.Genetic Variation: Some follicles
inherit weaknesses that make them more susceptible to atresia, contributing to
their higher numbers. However, in relation to this, there are several factors that
cause atresia, including hormonal imbalances, which refer to disruptions in
crucial hormones like FSH, LH, estrogen, and progesterone that can hinder
development and trigger atresia. Age and reproductive stage via ovarian
reserve naturally decline, especially during menopause.Environmental factors
such as toxins and medications can hurt their chances. Lastly, ovarian disorders
are conditions like PCOS that can disrupt development.

6. What are the main histological differences between spermatogonia and

spermatocytes?

● There are plenty of histological differences between

spermatogonia and spermatocytes?
● Spermatogonia:
- Location: Outer layer of seminiferous tubules
- Size: Small (10-15µm)
- Nucleus: Round, prominent nucleus
- Chromatin: Fine, dispersed chromatin
- Cytoplasm: Little cytoplasm
 - Arrangement: Single layer along the basement membrane
● Spermatocytes:
- Location: Deeper within the tubules
- Size: Larger (20-30µm)
- Nucleus: Irregular, reflects DNA increase during meiosis
- Chromatin: Condensed, forms bivalents/tetrads
- Cytoplasm: Abundant organelles for energy production
- Arrangement: Multiple layers within the tubules
● Therefore differences in structure and function distinguish
spermatogonia and spermatocytes during spermatogenesis.
Spermatogonia are the starting cells that divide mitotically to
produce spermatocytes. In contrast spermatocytes undergo
meiosis to create haploid spermatids which later develop into
sperm cells.

7. What is the main histological difference between spermatids and

spermatozoa?
One important histology-related difference between spermatids and
spermatozoa is their morphology. The process of spermiogenesis, which starts
with immature, spherical cells called spermatids, produces mature
spermatozoa. Spermatozoa are the fully developed, highly specialized cells that
have the ability to fertilize eggs. Moreover, Spermatids and spermatozoa vary
primarily in their histology; spermatids have spherical nuclei and no flagella,
whereas spermatozoa have elongated nuclei and flagella, which allow them to
move.

8. What structural features promote the fertilizing capacity of the sperms?

The structural features that promote the fertilizing capacity of the sperms
are the acrosome, mitochondria, and flagellum. The acrosome, which forms a
"cap" over most of the sperm cell's head, contains the lysosomal enzymes
needed to prepare sperm for the fertilization process. First, the mitochondria,
which is sperm’s midpiece that is incharge of producing ATP which help the
sperm to survive. In order for the flagellum to travel and reach the egg, the
sperm need ATP to accelerate their flagellum. Next, tail (flagellum), which
provides accessibility to the sperm to move towards the location of the egg.
Also, sperm motility and capacitation—the last stage of maturation needed for
fertilization—are maintained by a balance of ions that is regulated by a number
of ion channels and transporters found in the sperm plasma membrane.

9. Why are some seminiferous tubules devoid of cells?

Seminiferous tubules are the site where we find the production of
sperms. These are the tiny channels that can be found in the testicles. In
seminiferous tubules there is a cyclic process known as spermatogenesis which
includes the spermatogonia, meiosis and spermiogenesis; this cyclic process
mostly in adult testes may appear devoid of cells due to the cyclic nature of the
process. Due to the cyclic nature of spermatogenesis some tubules may not
contain cells at all stages, as spermatogonia constantly replenishes the
population. Also the pathological factors under certain circumstances, such as
inflammation, injury, or disease, the tubules may experience a reduction in the
number of cells due to cell loss or disruption.

10. Enumerate some of the functions of the Sertoli cells and Leydig
cells.
● Sertoli cells
Also known as the “nurse cell” which can be found located within the
seminiferous tubules of the testes that is responsible for spermatogenesis (the
process that generates sperm) in both humans and animals which provides
nourishment for the development and growth of sperms. In the seminiferous
epithelium, sertoli cells help growing germ cells by directly contributing to the
extracellular matrix's deposition and enabling the development of specialized
cell connections. Through phagocytosis, sertoli cells are in charge of
recognizing and eliminating aberrant, damaged, or apoptotic germ cells. The
developing spermatozoa's quality is preserved by this procedure. When sertoli
cells form strong connections with neighboring cells, the blood-testis barrier is
created. This barrier keeps harmful substances from the bloodstream from
getting inside the seminiferous tubules, protecting the developing germ cells.
Many hormones are secreted by sertoli cells, such as activin and inhibin,
which influence the manufacture of testosterone by Leydig cells and regulate
the hypothalamic-pituitary-gonadal axis.

● Leydig Cell
Also known as the “ interstitial cell” which can be found near the
seminiferous tubules that are responsible for the secretion of androgen.
Leydig cells release testosterone, the primary sex hormone in men. This
hormone has importance for several processes, such as the development of
secondary sexual features, the maintenance of germinal epithelium cells, and
the bodily control of sexual behavior. The pituitary gland secretes increased
LH during puberty, which activates Leydig cells. The development of
secondary sexual features, such as genital enlargement, facial hair growth,
muscle expansion, and voice deepening, is brought about by an increase in
LH encouraging Leydig cells to produce significant amounts of testosterone.
11. What histological features distinguish the frog's testis from the rat's
testis? Tabulate your answers.

Histological Frog's Testes Rat's Testes

Features
● Frogs have lobular ● The seminiferous
● Frog’s testis testicles, which are tubules in the
often made up of testis of rats are
many lobules linked and
consisting of convoluted in a
seminiferous tubular shape.
tubules encircled ● Rat testicular
● Bidder’s canal by interstitial tissue. spermatozoa are
● Spermatozoa in the frequently
seminiferous scattered along
tubules of the frog's the length of the
testis can often be seminiferous
found together in tubules instead of
grape-like clusters grouped or
or bundles. packed together.
● In frogs, the ovoid, ● Rats have bigger
white structures testicles than
that make up the frogs do. They
testes are paired are draped over
and situated the rump.
ventrally at the ● The ureter and
anterior end of the seminiferous
kidney. tubules are
● Sperm exit the frog connected by the
● Rat’s Testis testes through the rete testis, a
Bidder's canal, network of
which goes into the asymmetrical
kidney. tubules present in
● The frog's testicular rat testicles. The
tissue is lined with rete testis is a
densely packed crucial part of the
cuboidal cells. migration of
sperm.
12. Differentiate the sperms from the oocytes in terms of their morphology
and functions. Tabulate your answers.
Morphological and Functional Features
● Sperms - During sexual reproduction, the male
reproductive cell known as the sperm cell—also
called a gamete—is in charge of fertilizing the
female egg.
The general morphology of sperm cells
consists of the following parts:
- Head
- Midpiece (body)
- Tail

Functions:
● Head - contains the gamete's genetic material within its nucleus. A
structure known as the acrosome functions as a "cap" over the majority
of the sperm cell's head and is packed with lysosomal enzymes that are
necessary for getting sperm ready to engage in fertilization.
● Midpiece - The middle region of a sperm cell, situated between the head
and tail, is known as the midpiece. Similar to the head, the midpiece
accounts for roughly 10% of the entire length of the sperm. The midpiece
has densely packed mitochondria that generate the energy needed for
swimming.
● Tail - For the sperm, the tail is the source of motility. The migration of the
sperm toward the egg is caused by the tail's constant whipping motion.
● Oocytes - A female germ cell that can develop into an adult egg is
referred to as an oocyte. Also known as immature egg cells. The ovaries
contain oocytes, which are important for female reproduction.

● First polar body - At the beginning of the meiotic cycle, primary oocytes
generate the first polar body. Together with the little first polar body, the
primary oocyte also generates a larger secondary oocyte. A haploid initial
polar body and secondary oocyte are both present.
 ● Germinal Vesicle - The germinal vesicle (GV) is the nucleus of the
oocyte . The GV is large and spherical and contains chromatin (DNA)
and the nucleolus.
● Corona Radiata - a thick layer of tissue, surrounds the zona pellucida.
Proteins, carbohydrates, and hyaluronic acid make up this layer of cells.
Follicle cells stick to one another to form it. The hyaluronidase enzyme in
the sperm head facilitates the layer's breakdown, allowing the sperm to
access the egg. It also provides the ovum with the required proteins.
● Zona pellucida - the thick layer of glycoproteins known as the zona
pellucida envelops the plasma membrane of mammalian oocytes. A
crucial component of fertilization is the zona pellucida layer. The
acrosome reaction is started when the arriving spermatozoa unite with
the membrane. The adherence of the sperm membrane to the oocyte's
plasma membrane is known as the acrosome response. In humans, the
layer consists of cuboidal or columnar granulosa cells and is typically 13
μm thick. Sperm binding proteins are present in this layer, which
facilitates the sperm cells' effortless attachment to the plasma
membrane. It also keeps polyspermy from happening. As the embryo
approaches the uterine wall for implantation, the zona pellucida layer
sheds. Premature implantation (ectopic pregnancy) can be prevented by
the zona pellucida, which also plays a variety of functions in oocyte
development, spermatozoa binding, fertilization, protection during growth
and transport, and blastocyst development.
● Granulosa Cells - during the oocyte's development, these cells give it
support and nutrition.

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