EXERCISE 1: GAMETOGENESIS

Cells:
-

Somatic cells: give rise to all other cells in the body

Germ cells: give rise to gametes (egg and sperm)

Primordial germ cells (initial germ cells formed) -> developing gonads (where they divide mitotically) -> spermatogonia and oogonia (stem cells)
Stem cells undifferentiated cells that divide mitotically producing 2 types of cells
1. Remain as stem cells to keep cell line alive
2. Cells will become differentiated
- This nature that allows organisms to produce billions of sperms and eggs
Gametogenesis 1st phase in the sexual reproduction of animals
- process of formation of sex cells, which will laso undergo transformation to become specialized cells
- SPERMATOGENESIS (males) sperm formation
- OOGENESIS (females) ova formation
MAJOR PROCESSES:
1. Cell proliferation- primitive germ cells repeatedly divide by mitosis -> identical cells
2. Cell growth- cessation of mitosis; acquisition of raw materials for synthesis of protoplasm
3. Cell maturation- 2 successive cell divisions (meiosis) take place; accompanied by chromosomal division -> haploid number of
chromosomes
OVERVIEW:
Spermatogenesis:
- Round sperm > elongate > oval shape more pointed on one part
- Lose cytoplasm retaining only a small amount
- Form flagellum
Oogenesis:
- Do not transform much
- Increase in size and start to have a large amount of yolk
- Immotile
Common between them: They undergo MEIOSIS -> haploid # of chromosomes
*Embryo starts the development of organism and formed during zygote stage of an animal

SPERMATOGENESIS
-

Male gamete formation

Happens in the seminiferous tubules of the testis
Develops in association with specialized somatic cells
GRASSHOPPER

*slides were stained using Feulgen method

specific for DNA; chromosome and nuclear
structure; cytoplasm components- yellow/brown
-

devt takes place In the testis

each testis divided into lobes wherein blunt
ends facing the surgace of the organ; more
pointed ends directed towards the center, which
opens into the sperm duct (vas deferens)
Each lobe consists of many compartments
(cyst)
Each cyst contains clusters of similar cells
undergoing the SAME SOD
Cysts separated by septa made of CT
As devt proceeds, cysts pushed towards the
inner ends, and immature cysts take place at
the periphery
Growing cells are arranged from distal to
proximal
Blunt end: immature cysts
Pointed end continuous with the sperm duct:
mature and functional cysts

FROG
-

similar to grasshopper organization

wherein the seminiferous tubules of
the
testis is divided into cysts
active spermatogenesis restricted to
a short season in summer
sperm remains attached throughout
winter ( ~6 mos)
seminiferous tubules do not contain
permanent population of sperm cells
because they are evacuated at the
time of each spawning

MOUSE
*stained with H&E (blue: nuclei; pink:
cytoplasm)
* The same for CHICKS TESTIS but do
not stain clearly
- tunica albuginea: tissue
covering the testis; thick layer of
fibrous CT with a covering of
mesothelial cells
- subdivides the testis into
lobules, each containing
seminiferous tubules
- seminiferous tubules:
spherical; filled with
spermatogenic cells; convoluted
tubules with specialized
epithelium
- seminiferous epithelium made
up of sertoli cells and male sex
cells
- Spermatogenic cyce lasts a total
of 48 days
- Successive cycles start every 12
days
- 4 overlapping cycles

Sertoli/ Sustentacular cells:

- nurse cell
- supplies nutrients to spermatocysts
- associated with primary
spermatoginia and spermatocytes
as follicle cells
- at the time when tails of immature
sperms grow out and follicular cysts
burst open into the lumen, each
follicle cell attracts a bundle of
sperms and transforms into a
goblet-shaped nurse cell.

Sertoli cells
- spaced at fairly regular intervals
against the basement
membrane around the
circumference of the
seminiferous tubules
- near spermatogonia
- often surrounded by sperm
Functions:
1. form the blood-testes barrier
2. determine which substances
reach the germ cells
(spermatogonia are outside the
blood-testis barrier)
3. produce antigen-binding
proteins (necessary for
spermiogenesis)
2 types:
1. Active: pyramidal, apex towards
the lumen, and nucleus
perpendicular to tubule wall
2. Inactive: polygonal, nucleus
parallel to tubule wall

Spermatogonia:
- small, crowded; located at blunt ends
- nuclei: beaded appearance of chromatin; large
nuclei containing conspicuous masses of
chromatin
- These cells multiply for some time before
undergoing maturation

Spermatogonia:
- small, closely packed cells each with
granular oval nucleus
- each gonium invested by follicle
cells
- number of so-called residual
spermatozoid maintain this condition
permanently (source of germ cell
proliferation)

Spermatogonia:
- most primitive of the male sex
cells
- undergoes MD and their
progenies are translocated
(mediated by sertoli cells)
1. Rests on basement membrane;
interrupted by sertoli cells
2. Appears as thin layers of cells
with compact deeply staining
3. Nuclei tend to be quite small

Primary Spermatocyte:
- spermatogonium that has undergone growth;
ready to divide
- located near cysts containing sgonia
- very large nuclei
SUBSTAGES:
Interphase:
- inconspicuous, finely granular, may show a
single pale vascular nucleolus
Leptotene:
- chromatin crusts or granules -> finely beaded
filaments -> clearly filamentous
- chromosomes are at their longest (uncoiled),

Primary Spermatocyte:
- very large cells, irregularly spherical,
possess large and vesicular nuclei
- meiotic prophase configurations
may also be discerned

3 types of spermatogonia:
Type A:
- chromatin particles appear fine
and evenly dispersed
- single prominent chromatin
nucleolus, usually appears in an
eccentric position (these are
usually the germ line)
Type B:
- presence of coarse granules or
flakes of heavily stained
chromatin associated with the
nuclear membrane and
nucleolus
- most common, outnumbers tyoe
A and intermediate
- last stage before they transform
into primary spermatocytes
*most of the nuclei are in interphase
stage
INTERMEDIATE:
- with chromaphilic chromatin in
the form of delicate crusts or
flakes attached to the nuclear
membrane
- have prominent nuclei appearing
in a central position
Primary Spermatocyte:
- significantly large size and
nuclei
- chromosomes are very much
visible and the nuclear
membrane are not visible
(disintegrated)
Interphase:
- similar to gonial B
- Distinguished by finely granular
nucleus with pale nucleolus
- 3% total time of spermatogenic

appear to completely fill the cell

Zygotene/Synaptene:
- early stage: indistinguishable from leptotene
- thickened regions in the chromosomes
(indicates beginning of pairing [SYNAPSIS] of
homologous chromosomes
- synaptonemal complex: point of contact
between 2 homologs, assumes shape of long
loops attached by their extremity on 1 given
area of the nuclear envelop
- Configuration: Bouquet (drawn to one side)
- late stage: double thickness of chromosome
(advanced pairing)
Pachytene:
- chromosomes shorten and become thicker
- complete pairing and eventual exchange of
chromosomal arms happen [CROSSOVER]
- cells seem to be in haploid stage already
- stays like this for a long period of time; easily
identifiable
Diplotene:
- chromosomes partly split
- 4 strands in each homolog pair can be
discerned (due to relaxation and thickness)
- Chiasma first seen here and interchanges
between the homologs keep them united
- Configuration: X, cross, loop
Diakinesis:
- Coiling more evident here
- Chromosomes seen at shortest and thickest
- Configuration: loop, ring, bar, cross, etc.
Nucleus -> MI, AI, TI -> secondary spermatocytes
Secondary spermatocytes:
- smaller than primaries
- chromosomes not as evident
- stages of MDII may be seen

cycle (6 out of 212 hours)

Metaphase I:
- most easily recognized stage
- Male mouse: 20 discrete tetrads
are visible
- Tetrads are highly condensed,
especially deep staining, and
aggregated at the center part of
the cell (unlike in diakinesis)
Anaphase I:
- spindle usually distinct
- distinguished from metaphase I
by the visibility of spindle fibers
between the dyads
- Late Anaphase: shows cleavage
furrow
Telophase I:
- 2 groups of dyads that are
formed aggregate at the poles
- cytoplasm constricts to form 2
cells -> nuclear membranes
become distinct -> dyads uncoil
progressively

Secondary spermatocytes:
- half the size of primaries
- lie toward the lumen of the tubule
- darkly staining nucleus
- cytoplasm clearly visible and
prominent

Secondary spermatocytes:
- smaller than primaries
- may contain pale-stained
granular chromatin and several
globules of chromaphilic
chromatin free in the
nucleoplasm or associated with
the nuclear membrane

Spermatids:
- half the size of secondaries
- possess haploid set, but not functional as
gametes
- undergoes spermiogenesis: spermatids ->
sperm
Major events:
1. Spermatid elongates
2. Most of cytoplasm sloughed off, except for
small amount -> tail sheath (surrounds tail
which grows back from centrosome)
3. Chromatin mass becomes more condensed
and elongated to form solid staining head
Spermatozoa:
- ready to fertilize eggs
- consists of: head, middle piece (centrosome
and mitochondria), and tail
- long, thin, darkly staining heads
- abundant at distal region of the testis

twice the diameter of spermatids

and half as numerous

Spermatids:
- smaller than secondaries
- possess a condensed nucleus of
irregular shape
- cyst appears as cluster of granules
with dark nucleus being almost as
small as the cross section of the
sperm head
- inner of the two spermatid centrioles
passes into the nucleus while the
other one gives rise flagellum

Spermatids:
- smaller than spermatogonia
- initially quite round, pale staining
- nuclei -> elongate
- nucleus will decrease
- DNA will condense
- In early spermatids, the golgi
apparatus appears as an
intense red granule near the
nucleus
- Usually lie in close association
with sertoli cells during
spermiogenesis

Spermatozoa:
- 0.30mm
- elongated, solid staining head with
anterior acrosome pointing
outwardly toward the periphery of
the seminiferous tubule
- tail appears as a gray filamentous
extension of sperm head

Spermatozoa:
- deeply stained, short structures
- form the innermost layer of the
tubule
- heads of mature sperm are
buried in sertoli cells, tails
extend into lumen
- mature sperm: head, middle
piece, and tail
- head: circle-shaped and
flattened; tail: long and flagellate
Seminiferous tubules:
Between the seminiferous tubules, there
are:
a.) small blood vessels usually filled
with RBCs
b.) clump of cells conspicuous
nuclei that contains blue-stained
chromatin
c.) CT cells with elongated (spindleshaped) nuclei

Mesorchium:
- portion of peritoneum that suspends testis
to the dorsal body wall

Tunica albuginea:
- outer CT which covers the testis

known as intertubular CT or septula

extends into the substance of the
testis and partitions it into many
many compartments containing the
seminiferous tubules
Seminiferous tubules:
- much-coiled tube containing clusters
of spermatogenic cells undergoing
SAME SOD and nurse cells (sertoli
cells)
- connected with vas deferens
Spermatocyst/ Cell Nest/ Follicular cyst:
- cluster of similar cells undergoing
same SOD
- surrounded by follicle cells
- each cyst is formed from primary
spermatogonia and its associated
follicle
- at a certain stage, cyst wall breaks
down
- each seminiferous tubule may
contain cysts at different stages of
maturation
Interstitial cells:
- small cluster of cells found
interspersed with the tunica
albuginea
- secretes testosterone
Ova:
- egg cells seen between
seminiferous tubules of some male
individuals
- makes the male organ an ovotestis

OOGENESIS
-

Egg formation

Polar bodies: produced after 1st MD, smaller than egg due to unequal cytokinesis
This occurrence has the advantage of providing ovum with a much greater amount of cytoplasm and stored food
Polar body produced after 1st MD may or may not undergo 2nd MD; ALL POLAR BODIES ARE NON-FUNCTIONAL
Ootid differentiates quickly into ovum; doesnt have to undergo major changes
Oogenesis takes place within the follicles of the ovaries
Ovary covered by a continuous sheet of epithelium called the germinal epithelium (thought to give rise to future oocytes) but consists of
ovarian tissue
- Mesovarium: mesentery that attaches the ovary to the dorsal body wall; the route for blood vessels to and from the ovary
- Ovary divided into:
1. Central medulla
- consists largely of CT and blood vessels; continuous with mesovarium
2. Peripheral cortex
- business part
FROG
CAT
HUMAN
- essentially a large yolk of sac with
Germinal epithelium:
heavier and larger granules
- epithelial lining of ovary composed of
concentrated toward the animal
cuboidal cells
hemisphere and in the vicinity of
Tunica albuginea:
germinal vesicle (immature nucleus)
- fibrous CT beneath germinal epithelium
- Coat: surrounds entire egg; necessary
Stroma:
for retaining shape and aids in
- CT beneath tunica albuginea
cleavage and gastrulation; contains
- Penetrates the substance of the ovary
pigment (metabolic by-product)
- Consists of CT fibers and spindle
shaped cells
Mesovarium:
Corpus luteum
- double-layered extension of peritoneum;
- may be in various stages of regression
attaches ovary to the dorsal body wall
- large cavity filled with loose CT and
remnants of lutein cells
Ovarian follicle:
Blood vessels
-collective term for theca externa, theca
- vein and artery
interna, and follicle cells
-

Theca externa:
- outermost CT surrounding ovary
- continuous with mesovarium
- region of rupture during ovulation to
allow egg to escape into body cavity
- thousands of individual sacs
containing eggs are suspended in this

covering
Theca interna:
- inner CT surrounding ovary and each
egg
*Theca externa and interna contain blood
vessels and nerves. They make up the limited
stroma of the frogs ovary
Follicle cells:
- cells derived from the oogonia
- contain oval and granular nuclei,
surrounds the developing oocyte
- found in close association with the
oocyte through maturation
Oogonia:
- usually located at the periphery of
ovary; much smaller than oocytes
- no pigment and contain little/no yolk

Both follicle cells and ovum come from

original oogonia
Undergoes mitosis -> growth: Ova
Fail to grow: follicle cells

SoM: PRIMARY FOLLICLE STAGE

immature egg cell becomes
surrounded by a single layer of
flattened follicular cells
Structures present:
a. Follicles
- single layer of cells surrounding
oogonium
- have dark nuclei and may be
squamous or low cuboidal
b. SoD: Oogonium
- surrounded by single layer of cells
- bigger than follicle cells
- spheroidal and has vesicular nucleus
with prominent nucleolus
- embedded in the stroma and occurring
in clusters
*In the late primary follicle stage:
Egg cell moves deeper into ovary -> more
mature and starts to enlarge -> separated from
follicle cells that surround it by zona pellucida
-> follicle cells proliferate mitotically -> multiple
layers (growing follicle)
* the stromal cells adjacent to the follicle form
a specialized sheath or theca around it
-

Tunica albuginea and stroma:

- more prominent in humans

SoM: GROWING FOLLICLE STAGE

follicle cells several layers thick
increased in size and complexity like
egg cell
- antrum, zona pellucida, stratum
granulosum start to be formed
Structures present:
a. Follicles
- several layers thick consisting of low
columnar or cuboidal cells
b. Antrum
- follicular cavity
- formed by union of spaces between
follicle cells (may not be evident)
- granular appearance of cavity due to
precipitation of protein in the fluid
c. Liquor folliculi
- viscous fluid secreted by follicle cells
- appears as light pink coagulum within
antrum
d. SoD: Oogonium/ Primary oocyte
- separated form the granulosa cell by
the zona pellucida
e. Zona pellucida
- deeply staining membrane
surrounding the egg cell, separating it
from follicle cells
- secreted by germ cell and the follicle
cells together
*At the onset of antrum formation,
- Oocyte reach its max size but follicle
continues to grow
- Follicle (granulose) layer continues to
proliferate -> theca externa and interna
- Spaces filled with liquor folliculi form in
the stratum granulosum
- Irregular spaces then join forming
antrum
-

SoM: GRAAFIAN FOLLICLE STAGE

- large follicles that bulge on the free
surface of ovary
Structures present:
a. Antrum
- at this point, it is bigger
- lined by granulose cells, stratum
granulosum, which is a thick layer of
follicle cells
b. Cumulus oophorus
- eccentric mound of follicle cells
surrounding egg cells
- includes oocyte, zona pellucida, and
surrounding granulose cells
c. SoD: Primary oocyte
- heavy nuclear envelop
- granular nucleus with large nucleolus
and yolk-filled cytoplasm
d. Stratum granulosum
- layers of follicle cells bordering
Graafian, excluding cumulus oophorus
e. Corona radiata
- layer of follicle cells of cumulus
oophorus lie nearest to oocyte
- separated from oocyte by zona
pellucida
f. Zona pellucida and Liquor folliculi
g. Theca folliculi
- stroma immediately adjacent to the
follicle that form a capsule around the
follicle
g.1. Theca interna (thicker)
- inner layer between theca externa and
granulose cells of capsule
- consists of large stromal cells with oval
nuclei and capillaries
- richly vascularized
- secretes estrogen
g.2. Theca externa (thinner)
- outer layer of capsule composed of

spindle-shaped dense CT fibers

continuous with the surrounding
ovarian stroma
*Ovulation will involve rupture of ovarian
surface and follicle wall
-

h. Atretic follicles
- unsuccessful follicles undergoing
degenerative process
*Initial atresia- occurs in the ovum after in
which there is degeneration of follicle cells
Resulting structure: scattered follicle cells,
unorganized stratum granulosum, and germ
cell fragments
- Atresia of growing follicle: presence of
collapsed ZP and cavity invaded by theca
interna (corpus atretica)
i.
-

Corpus lutea
masses of granular tissue in the ovary
resulting from ovulation of egg cells
from follicles
- persist during pregnancy; responsible
for production of progesterone
- lightly stained and follicles seem to be
converging at the mid portion
Structures present in corpus luteum:
1. Lutein cells
- Granulosa lutein: large, light stained,
from stratum granulosum, produces
progesterone
- Theca lutein: smaller, dark stained,
from theca interna, produces estrogen
and progesterone
j. Capillaries
- network of small blood vessels with a
single endothelial lining
- identified by blank spaces or by dark
masses of blood corpuscles
k. Capsule

layer surrounding the corpus luteum

consists of CT fibers and fusiform cells
(both derived from theca externa),
lutein-like cells (from theca interna),
and blood vessels
l. Corpus albicans
- found in the central portion of ovary
- non-functional corpus luteum formed
when fertilization does not take
place
- stroma forms a capsule around the
regressing corpus luteum, invades
regressing luteal cells and replaces
them
- leaves a white hyalinized scar (pale in
central portion)
m. periovarial membrane
- peritoneum of ovary which continues
into the oviduct
- consists of squamous epithelial cells
and adipocytes
n. Periovarial space
- space between periovarial membrane
and the ovary
o. Oviduct
- convoluted structure alongside the
ovary
- attached to periovarial membrane
Oocytes:
- bigger than oogonia
- germinal vesicle (immature nucleus):
lightly stained at the center
- referred simply as oocytes (primary
and secondary cannot be
distinguished)
Vitelline membrane:
- non-cellular, transparent membrane
between follicle cells and each mature
egg

derived from both ovum and follicle

cells
- developed during maturation stage,;
not seen in early oogonia stage
Perivitelline membrane:
- fluid-filled space formed between egg
and vitellline membrane AFTER
fertilization
- fluid may be derived from egg; shows
compensatory shrinkage