~ Grasshoppers Testis ~ - Paired chromosome assumes the shape of long loops
attached by their extremity on one given area of the
Development of the spermatozoa, takes place in the male gonads.
Each testis is divided into several loves makes the grasshoppers
testis unique among the other testis
Cyst
- Separate compartments inside the lobe
- Contains clusters of similar cells undergoing the same
stage of development
- Each cysts are separated from each other by means of
septa (composed of connective tissues)
As development proceeds the cysts is pushed towards the inner of
the lobe, and more immature takes its place at the periphery.
Fuelgen method
- Specific for DNA
- Useful for studying chromosome and nuclear structure
- Cytoplasmic components are stained yellow or brown
SPERMATOGONIA
- Cells are small, crowded
- Located at the blunt end of the testis
- Nuclei show a beaded appearance of the chromatin
material
- Have a large nuclei containing conspicuous masses of
chromatin
-
PRIMARY SPERMATOCYTES
- Spermatogonium that has undergone growth
- Located in the cysts nearest those containing
spermatogonia
- Have a very large nuclei
INTERPHASE
- Inconspicuous spermatocytic interphase that appears
finely granular
- May show a single pale vascular nucleolus
LEPTOTENE
- Chromatin crusts or granules resolve into finely beaded
filaments
- Chromatin becomes clearly filamentous
- Chromosomes are at their longest (uncoiled stage), they
appear to completely fill the cell
ZYGOTENE
- Have thickened regions in the chromosomes which
indicate the beginning of pairing (synapsis) of homologous
chromosomes
- Synaptonemal complex point of contact between two
homologues
nuclear envelope
- Late zygotene double thickness of the chromosome
strands indicate advanced pairing
PACHYTENE
- Chromosomes shorten and become thicker
- In this stage, complete pairing and eventual exchange of
chromosomal arms happen.
- Cells seem to be in haploid condition already
- The chromatin stays in this condition for a long period of
time, thus pachytene is very easy to identify. In some cases,
a lobe is half-filled with cyst.
DIPLOTENE
- Chromosomes partly split
- Four strands in each homologous pair can be discerned
due to relaxation of pairing
- Chromosomes are thicker in appearance
- Chiasma is first seen, interchanges between the
homologues keep them united
- Configurations like an X or a cross or a loop may be
observed
DIAKINESIS
- Progressive condensation of chromosomes by means of
coiling is more evident
- Chromosomes are seen at their shortest and thickest
- Each pair shows various configurations (loop, ring, cross,
bar) depending on the position and degree of
terminalization of one or more chiasmata.
SECONDARY SPERMATOCYTES
- Smaller than the primaries
- Chromosomes are not evident
SPERMATIDS
- Spermatids first resemble ordinary cells about half of the
size of secondary spermatocytes
- Possess a haploid set of chromosomes but are still not
capable of functioning as male gametes
- ^ have to undergo spermiogenesis (transformation of
spermatids to spermatozoa and can be seen in the inner
regions of the lobe)
- The major events that happened during this period involve
major nuclear and cytoplasmic changes that eventually will
terminate in the formation of the functional sperm.
- Spermatid elongates, most of the cytoplasm is sloughed
off, except for a small amount which becomes that tail
sheath surrounding a fine tail filament which grows back
from the centrosome.
- Chromatin mass becomes more condensed and elongated
to form the solidly staining head, middle piece containing
the centrosome and mitochondria, and the extremely
long tail.
SPERMATOZOA
- Mature sperm cells
- Long, thin and darkly staining heads with distinct tails
which may measure seven to eight times longer than the
heads
- Abundant at the distal region
~ Frogs Testis ~
Mesorchium
- portion of the peritoneum that suspends the testis to the
dorsal body wall
- immediately ventral to the kidney
- Between the layers of which pass the tubes of the vas
deferens
Tunica albuginea
- Outer connective tissue which covers the testis
- Known as intertubular connective or septula
- This tough tissue extends into the substance of testis and
partitions it into many compartments containing the
seminiferous tubules
Seminiferous tubule
- A much coiled tubule containing the clusters of
spermatogenic cells undergoing the same stage of
development and nurse cell, sertoli cells.
- Connected with the vas deferens
Spermatocyst/Cell nest/Follicular cyst
- A cluster of similar cells undergoing the same stage of
development and is surrounded by follicle cells.
- Each spermatocyst is formed from a primary
spermatogonium, and its associate follicle and forms
gonia of the second and third order, which may go
maturation divisions and forms spermatozoa.
- At a certain stage, the cyst wall breaks down.
SPERMATOGONIA
- Small, closely packed cells, each with a granular oval
nucleus
- Primary spermatozoa are fasted to the walls of the
seminiferous tubule
- Each gonium invested by follicle cells
PRIMARY SPERMATOCYTES
- Very large cells that tend to be irregularly spherical,
possessing large and vesicular nuclei
SECONDARY SPERMATOCYTES
- Cells that develop as the result of the first meiotic division
that is about half the size of the primaries
- Lie towards the lumen of the tubule
- Generally have a darkly staining nucleus
- Cytoplasm clearly visible and prominent
SPERMATIDS
- Smaller than the secondary spermatocytes
- Possess a condensed nucleus of irregular shape
- Cluster of metamorphosing spermatids appear as a
cluster of granules with dark nucleus
- inner of the two spermatid centrioles passes into the
nucleus while the other one gives rise to the tail-like
flagellum
SPERMATOZOA
- cells average 0.30mm
- each sperm has an elongated, solid staining head (nucleus)
with an anterior acrosome (pointing outwardly toward the
periphery of the seminiferous tubule) tail appears as a
gray filamentous extension of the sperm head
Sertoli or sustentacular cell
- Goblet-shaped
- Nurse cell, supplying nutrient to the cluster of cell until
such time when they may be liberated into the lumen and
through the genital tract to the outside
- Early associated with the primary spermatogonia and
spermatocytes as spermatocytes
Interstitial cells
- small clusters of cells found interspersed with the
intertubular connective tissue
Ova
- Egg cells seen between seminiferous tubules of some male
individuals, thus making the male organ an ovotestis.
~ MOUSES TESTIS ~
Stained with hematoxylin (blue stain which stain the nuclei blue)
and eosin (pinkish-orange counterstain that is concentrated by
acidic components of the cells including the cytoplasm)
Covering the mouse testis is the tunica albuginea which is a thick
layer of fibrous connective tissue with a covering of mesolecithal
cells. This connective tissue subdivides the testis into a number of
lobules, each containing seminiferous tubules.
The seminiferous tubules are filled with spermatogenic cells, and
they are made up of specialized seminiferous epithelium (consists of
Sertoli cells (somatic cells) and male sex cells undergoing
spermatogenesis and spermiogenesis.
Spermatogenic cycle in Mice last a total of 48 days but successive Nuclei tend to be quite small, comparable in size
cycle start every 12 days so that there are a total of 4 overlapping to those of spermatids
cycles at every site along the tubule. - Three types of spermatogonia:
Type A Spermatogonia
Seminiferous tubules have
o Chromatin particles appear fine and evenly
- Small blood vessels that are usually filled with red-stained
dispersed
erythrocytes
o A single prominent chromatin nucleolus, whose
- Clump of cells with conspicuous nuclei that contains
nature is not known, usually appears in an
clumps of blue-stained chromatin
eccentric position.
- Connective tissue cells with elongated (spindle-shaped)
o These are usually the germ cell line
nuclei
Type B Spermatogonia
o Chromatin is identified by the presence of coarse
granules or flakes of heavily stained chromatin
Interstitial cells (Leydig cells)
associated with the nuclear membrane and
- Cells with conspicuous cells
nucleolus
- Produce testosterone
o Most common gonia
o Last stage before they transform into primary
spermatocytes
Sertoli cells
- Subtentacular cells which are spaced at fairly regular Although occasional metaphase and anaphase are seen in this layer,
intervals against the basement membrane and around the most of the nuclei are in the interphase stage, characterized by the
circumference of the seminiferous tubules, therefore near presence of irregular blocks of chromatin.
the spermatogonia
Intermediate spermatogonia
- Have distinct outlines and large, oval, often indented
- Spermatogonia with chromophilic chromatin in the form
nuclei with compound nucleoli.
of delicate crusts or flakes attached to the nuclear
- Often surrounded by spermatozoa
membrane
- As supporting cells:
- Have prominent nucleoili usually appearing in a central
Form the blood testis barrier: nutrients and
position rather than peripheral position
circulating substances do not directly reach the
germ cells
Determine which substances reach the germ
Primary Spermatocytes
cells. The spermatogonia are outside the blood-
- Large in size
testis barrier
- nuclei are large than the spermatogonia
Produce antigen biding proteins, which are
- The only other large nuclei within the tubles belong to
necessary for spermiogenesis.
Sertoli cells and these even have a different morphology
- Two types of sertoli cells:
and lie at a different level
Active sertoli cells
- Chromosomes are very much visible and the nuclear
o Cell which is fulfilling its normal function of
membrance, are disintegrated already and therefore not
supporting spermatozoa
visible anymore.
o Pyramidal in shaoe
o Apex directed towards the lumen
o Nucleus perpendicular to the tubule wall
Interphase
Inactive sertoli cells
- Similar to gonial B in appearance but distinguished from it
o Polygonal in shape
principally by the finely granular nucleus with its pale
o Nucleus is parallel to the wall of the tubule
nucleolus
Translocation is mediated by Sertoli cells - Very short in duration
- Griffen and MacMahon have estimated the time of
Spermatogonia
duration for this stage to be approximately 3% of the total
- Most primitive of the male sex cells
time of spermatogenic cycle as determined by Oakberg.
- Some spermaotogonia undergo meiosis and their
- Interphase exist for about 6 hours
progenies are translocated, as they mature, in groups
around the circumference of the seminal tubule toward the
Metaphase I
lumen.
- 20 discrete tetrads are visible
- Three criteria to identify spermatogonia:
- Tetrads are highly condensed, especially deep staining and
The gonia rests on the basement membrane
aggregated at the center part of the cell.
and interrupted at intervals by sertoli cells
The gonial layer, owing to their rapid mitotic
activitiy, appear as thin layers of cells with
Anaphase I
compact deeply staining nuclei forming a poorly
- Spindle is usually quite distinct
defined row immediately internal to the wall of
- Visibility of spindle fibers between the dyads
the seminiferous tubule
- Late anaphase may show cleavage furrow already OOGENESIS
- Process of producing egg cells. The eggs are products of
meiotic division, which is responsible for the haploid
Telophase I
condition of the cells.
- After Anaphase I, two groups of dyads that formed are
- A polar body is produced after the first meiotic division.
aggregated at the poles.
- Polar bodies are very much smaller than the egg due to
- As the cytoplasm constricts to from the two cells, nuclear
unequal cytokinesis that occurs.
membranes become distinct and the dyads uncoil
- The occurrence has the advantage of providing the ovum
progressively to from the second meiotic interphase.
with a much greater amount of cytoplasm and store food
than if an equal division were to occur.
- The polar body produced after MD1 may or may not
Secondary Spermatocytes
undergo a 2nd meiotic division depending on the condition
- Spermatogenic cells resulting from the first meiotic
of the cell.
division of the primary spermatocyte
- All the polar bodies are non-functional.
- Small than the primaries
- Unlike sperm cells, an ootid doesnt have to go through
- May contain a pale stained granular chromatin, several
major changes for it to become a functional ovum.
globules of chromophilic chromatin free in the
- In fact after meiosis, the ootid quickly differentiates into
nucleoplasm or associated with the nuclear membrane
mature egg cell or ovum.
- They grossly resemble spermatids with which they
- Takes place within the follicles of the ovaries
regularly have been confused; however, they are twice the
- Ovary is covered by a continuous sheet of epithelium
diameter of spermatids and half as numerous
called the germinal epithelium (consists of the ovarian
tissue not the future egg cells)

Spermatids
Mesovarium
- Result of the Meiotic II
- Mesentery that attached the ovary to the body wall
- Lie next inward beyond the spermatocyte later in the
- Route for blood vessels to and from the ovary
seminiferous tubules
- Sizes approximate that of spermatozoid but still smaller
- Spermatid nuclei are quite round; abundance, small size,
- The ovary itself is divided into the central medulla and a
pale staining properties (they are haploid) and position
more peripheral cortex.
above the level of primary spermatocytes.
- The medulla consists large of connective tissue and blood
- Nuclei dont remain round for long they progressively
vessels and appears more or less continuous with the
elongate; their DNA soon undergoes a progressive
mesovarium.
condensation
- The cortex is the business part of the ovary.
- Nucleus will progressively decrease in size due to the
elimination of nuclear sap, and increase in staining
intensity as the DNA becomes more and more condensed. Ovary
- In early spermatids, the golgi apparatus appears as an
intense red granule near the nucleus. Central
Cortex
Medulla
- The flagellum appears relatively early and in later stages,
the midpiece is represented as a pink-stained thickening connective business part
along about the first half of its length. tissue of the ovary

- Spermatids usually lie in close association with sertoli cells

during their transformation into spermatozoa. blood vessels

Spermatozoa ~ Frogs Ovary ~

- Mature spermatozoa appear in sections as deeply stained,
short structures, which form the innermost layer of
spermatogenic tubule.
- The heads of mature spermatozoa are buried in sertoli
cells, and the tails extend into the lumen.
- The fully mature spermatozoa becomes free of the nurse
cells and pass into the ducts.
- Found in the lumina of the testis tubules and ducts, made
up of head, middle piece and tail.
- Head is circle shaped and flattened, and the tail long and
flagellate.
- Essentially a large sac of yolk with the heavier and larger
granules concentrated toward the animal hemisphere and
in the vicinity of the germinal vesicle or immature nucles
- Surrounding the entire egg is a coat necessary for retaining
the shape of the egg and aids in cleavage and gastrulation.
- This coat contains pigment which is a metabolic by-
product.
Mesovarium
- A double-layered extension of the peritoneum
- Attaches the ovary to the dorsal body wall
 Ovarian follicle
- Collective term for theca externa, theca interna and the
follicle cells
Theca externa
- Outermost connective tissue surrounding the ovary.
- Continuous with the mesovarium
- Region of rupture during the ovulation to allow the egg to
escape into the body cavity.
- Thousands of individual sacs containing the eggs are
suspended in this covering.
Theca interna
- Inner connective tissue surrounding the ovary
Theca externa and Theca interna
- Contain blood vessels and nerves
- Make up the limited stroma of the frogs ovary
Follicle cells
- Cells derived from the oogonia
- Contain oval and granular nuclei
- Surrounding the developing oocyte
Oogonia
- Usually located at the periphery of the ovary and much
smaller than the oocytes.
- Have no pigment and contain little, if any, yolk.
Each cluster of oogonia, representes a future ovarian unit, consisting
of many cells and one ovum.
Both follicle cells and ovum come from original oogonia.
These pre-maturation germ cell divides by mitosis numerous times
and then enters a quiescent phase, during which there is growth.
These become ova while those that fail become follicle cells.
The majority of oogonia, therefore, never mature into ova.
Oocytes
- Potential ova that is bigger than the oogonia
- Lightly stained at its center is the germinal vesicle or
immature nucleus
Vitelline membrane
- Non-cellular, transparent membrane between the follicle
cells and each mature egg.
Perivitelline space
- A fluid-filled space from between the egg and the vitelline
membrane
- Fluid maybe derived from the egg, which shows
compensatory shrinkage
~ Cats Ovary ~
Germinal epithelium
- Epithelial living of the ovary that is composed of cuboidal
cells
Tunica albuginea
- A fibrous connective tissue beneath the germinal
epithelium
Stroma
- Connective tissue immediately beneath the tunica
albuginea and penetrating the substance of the ovary
- Consisting of connective tissue fibers and spindle shaped
cells
Oogonium
- A potential ovum with vesicular nucleus with a prominent
nucleolus.
- Found embedded in the stroma and occurring in clusters
- Small oogonia without the follicle cells surrounding them
are in their early stage
Primary follicle stage
- Immature egg cell that is embedded in the stroma now
becomes surrounded by a single layer of flattened
follicular cells.
- A primary follicle consists of:
Follicles
o Single layer of cells that surround the developing
germ cell.
o Have a dark nuclei and may be flattened
(squamosal) or cuboidal, depending on the
developmental stage of the follicle
Oogonium
- A potential ovum surrounded by a single layer of follicle
cells.
- Bigger than the follicle cells surrounding it
- Spheroidal and has a vesicular nucleus with a prominent
nucleolus
In the late primary follicle stage, the immature egg cell, together
with its surrounding layer of follicle cells, start to move deeper in the
ovary. The egg cell is now more mature and starts to enlarge.
It is then separated from follicle cells that surround it by the zona
pellucida.
The follicle cells, which are initially in a single layer, now proliferate
mitotically to form multiple layers now called growing follicle.
The stromal cells adjacent to the follicle form a specialized sheath or
theca around it.
 Growing follicle stage
- Immediately succeeds the primary follicle stage
- During this stage, the follicle cells have proliferated by
dividing mitotically and are now several layers thick.
- Has increased in size and complexity like the egg cell
- The antrum, zona pellucida and stratum granulosum start
to be from during this stage.
- The following structures make up a growing follicle:
Follicle cells
o Several layers thick and consisting of low
columnar of cuboidal cells enclosing the oocyte
Antrum
o Also known as the follicular cavity
o Cavity within the growing follicle and is formed
by the union of spaces between the follicle cells
o Precipitation of protein in the fluid may give the
cavity a granular appearance
Liquor folliculi
o A viscous follicular fluid secreted by the follicle
cells
o Appears as a light pink coagulum within the
antrum
Oogonium or Primary oocyte
o The probable sage of development of the cell in
a growing follicle
o During this stage, the oogonium/primary oocyte
is separated from the granulosa cell by the zona
pellucida
Zona pellucida
o A deeply staining membrane surrounding the
egg cell
o Separates it from the follicle cells
o Secreted by the germ cell and the follicle cells
An the onset of antrum formation, the oocyte has actually reached it
maximum size but the follicle as a whole continuous to grow in size
until it becomes a large vesicle, which bulges from the free surface of
the ovary.
The follicle cell layer continuous to proliferate, the theca interna, a
vascular inner layer of estrogen-producing secretory cells and the
theca externa composed of primarily of connective tissue.
As this continued to progress, spaces, filled with liquor folliculi, are
formed in the stratum granulosum.
The irregular spaces then join forming a large fluid-filledcavity
known as the follicular cavity or antrum.
Graafian follicles
- Large follicle that bulges on the free surface of the ovary
Antrum
- Fluid filled cavity
- First seen in the growing follicle
- Only this time, it has grown bigger
- The antrum, at this point, is lined by granulose cells
(stratum granulosum) thick layer of follicle cells
Cumulus oophorus
- The eccentric mound of follicle cells surrounding egg cells,
it includes the oocyte, zona pellucida and surrounding
granulose cells.
Primary oocyte
- Has a heavy nuclear envelope, a granular nucleus with a
large nucleolus and a cytoplasm filled with yolk granules
It may not be possible to see a secondary oocyte in a section of the
ovary because ovulation occurs immediately after MI. The primary
oocyte becomes a secondary oocyte after the first maturation
division.
Stratum granulosum
- Layers of follicle cells bordering the graafian, excluding the
cumulus oophorus
Corona radiate
- Layer of follicle cells of the cumulus oophorus that lie
nearest to the oocyte
- Separated from the oocyte by the zona pellucida
Theca folliculi
- The stroma immediately adjacent to the follicle that forms
a capsule around the follicle
Theca interna
- Inner layer between the theca externa and the granulose
cells of the capsule consisting of large stromal cells with
oval nuclei and capillaries
- Richly vascularized
- Cells are secretory
Theca externa
- Outer layer of the capsule composed of the spindled-
shaped dense connective tissue fibers.
- Continuous with the surrounding ovarian stroma
- Arranged as a more or less discrete capsule
Ovulation will involve the rupture of the ovarian surface as well as
the wall of the follicle.
Atretic follicles
- Unsuccessful follicles undergoing degenerative process
Initial atresia occurs in the ovum after in which there is degeneration
of follicle cells.
The resulting structure shows an antrum with scattered follicle cells,
an unorganized stratum granulosum and fragments of the germ
cells.
Atresia of a growing follicle is exhibited by the presence of a
collapsed zona pellucida and a cavity invaded by the thickened and
vascularized theca interna.
The mass of tissue resembles a regressing corpus luteum and is
called corpus atretica.
 Corpus lutea
- Masses of granular tissue in the ovary resulting from the
ovulation of egg cells from follicles.
- Persist during pregnancy and are responsible for the
production of progesterone.
- More lightly stained and the follicles seem to be
converging at the middle portion
- Structures in a corpus luteum:
Lutein cells
o Large, pale staining cells with large vesicular
nuclei
o Cells come from the stratum granulosum of the
ovulated graafian follicle.
o Located on the periphery of the corpus luerum
and within the contours of its folds
Capillaries
o Network of small blood vessels with a single
endothelial lining
o Identified usually as empty spaces or by dark
masses of blood corpuscles
Capsules
o Layer surrounding the corpus luteum, consisting
of connective tissue fibers and fusiform cells
(both derived from theca externa), lutein-like
cells (from theca interna) and blood vessels
Corpora lutea
- Most of the corpora may be in various stages of regression
thus an active corpus luteum may not be seen
The degenerating corpus lutea in human, unlike those of the cat, has
a large cavity filled with loose connective tissue and remants of
lutein cells.
Blood vessels
At ovulation, the follicular wall will burst open and the ovum,
together with its surrounding cumulus cells and liquid contents of the
follicular cavity, will be cast out into the oviduct or fallopian tube.
The first meiotic division happens shortly before ovulation.
The secondary oocyte is released at metaphase of the second meiotic
division; it and the first polar body are still contained within the zona
pellucida. The latter is surrounded by a halo of granulosa cells, which
are collectively called the zona radiate (in some texts the adherent
granulosa cells are said to comprise both the zona radiate and a
cumulus oophorus layer. This composite group of cells and the
follicular fluid are released at ovulation. It is of interest that not all
the follicles that can be found In the ovary reach the point of
expelling ripe eggs. Most of them break down and are resorbed at
some stage of their development; many of the primordial follicles
never develop any further.

Corpus albicans
- Usually found in the central portion of the ovary
- Non-functional corpus luteum is formed when fertilization
does not take place
- Stroma forms a capsule around the regressing corpus
luteum, invades the regressing luteal cells and replaces
them as they degenerate leaving a white hyalizined scar
- Identified by the pale stain in its central portion

Periovarial membrane
- Peritoneum of the ovary which continues into the oviduct
- Consist of squamous epithelial cell and adipocytes

Periovarial space
- Space between the periovarial membrane and the ovary

Ovary
- Convoluted structure that is found alongside of the ovary
and is attached to the periovarial membrane

~ Human Ovary ~
- Similar with cats ovary

Tunica albuginea and stroma

- More prominent in humans, being bigger