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HISTOLOGY OF FROG TESTIS

No arrangement/organization

Structures
1. Mesorchium
a. Peritoneum that suspends testis to dorsal
body wall
Mesorchium
b. Between layers of tubes of vas deferens
2. Tunica Albuginea
a. Outer connective tissue
b. Covers testis
c. Known as intertubular connective tissue or
Seminifer
septula
ous
d. Tough tissue; extends into substance of testis
e. Partitions into many compartments containing
Tubules
seminiferous tubules
Tunica Albuginea
3. Seminiferous Tubules
a. Coiled tubule containing spermatogenic cells
b. Also has sertoli cells nurse cells; nutritional;
going to gonadal layer
c. Tubule connected to vas deferens
d. Spermatogenic cells may undergo
maturation
Primary
e. Cells going through a particular stage found
Spermatocyte
in a culture/group within the tubule
4. Spermatocyst or Cell Nest or Follicular Cyst
a. Cluster of similar cells undergoing same
stage of devt
Cysts of
b. Surrounded by follicle cells
Spermatogonia
c. Spermatocyst = primary spermatogonium +
associate follicle
Secondary
d. Spermatocyst forms gonia of the second
Spermatocyte
and third order which undergo maturation
to form SPERMATOZOA
e. Cyst wall eventually breaks down
f. Spermatocysts mature independently of
one another
Spermatogo
g. Each seminiferous tubule may contain
nia
Spermatoz
cysts in different stages of maturation
oa
5. Spermatogonia
a. Closely packed cells with granular nucleus
b. Heavily stained; not a perfect circle, different shapes
c. Primary spermatozoa fastened to walls of seminiferous tubule
d. Gonium invested by follicle cells
e. Residual spermatozoid source of germ cell proliferation
f. Active spermatogenesis
i. Short season in summer
ii. Begins rapid succession of mitosis
iii. Daughter cells from spermatogonial divisions retain follicle cells
1. Eventually become sperms
g. Millions of sperms from frog come from small number of gonia by repeated mitosis, maturation
and spermatogenesis

Secondary Spermatocyte

Spermatogonia

h. Seminiferous tubules
i. Do not contain permanent population of sperm cells
ii. Sperm cells evacuated at time of each spawning
6. Primary Spermatocytes (6,7,8 early spermatids perfect circles)
a. Very large cells
b. Irregular, large and vesicular nuclei
c. Meiotic prophase can be discerned
d. Divides to form haploid secondary spermatocytes
e. Middle not stained
f. You should see chromosomes; biggest
Primary Spermatocytes
7. Secondary Spermatocytes
a. Haploid; result of first meiotic division
b. Half the size of primaries
Spermati
c. Stain half; darkly stained but white at sides
d
Spermato
(cytoplasm)
d. Lie toward lumen of tubule
zoa
e. Clearly visible cytoplasm
8. Spermatids
a. Condensed nucleus
b. Closely packed you shouldnt see any white
c. Cluster of granules
d. Metamorphosis of spermatids to spermatozoon
i. Inner of 2 spermatid centrioles passes into nucleus while the other one gives rise to
the tail-like flagellum
9. Spermatozoa
a. Average at 0.30 mm
b. Elongated solid staining head
c. Anterior acrosome pointing outward to seminiferous tubule
d. Tail is a gray filamentous extension of head
e. Groups into 25 to 40 cells for external sources of nutrition
f. Columnar type basal cell known as sertoli cells
i. Sertoli cells; source of nutrition for sperm heads
10. Sertoli or Sustentacular Cell
a. Nurse cell; supplying nutrients to sperm
b. Was associated with the primary spermatogonia and spermatocytes as follicle cells
c. When the tails of immature sperms grow out and follicular cysts (spermatocysts burst open
into lumen of seminiferous tubule)
i. Each bursted follicle cell attracts a bundle of sperm and transforms into a goblet
shape nurse cell
d. Frog sperms attach through winter, as long as 6 months
i. When detached, theyre stored in the mesonephric tubules and in different ducts
ii. In the mesonephric tubules they die agad
11. Interstitial Cells
a. Small clusters of cells interspersed with intertubular connective tissue
b. Undetermined if these cells secrete male sex hormone in the frog
12. Ova
a. Egg cells seen between seminiferous tubules of some male individuals
b. Male organ OVITESTIS
HISTOLOGY OF MOUSE TESTIS (chick testis also applicable here)

Arranged from basement membrane to lumen


Stained with hematoxylin and eosin
o Hematoxylin
Blue stain that stains nuclei
o Eosin
Pinkish-orange counter stain to contract
acidic components
Cytoplasm
Tunica albuginea
o Tissue covering mouse testis
o Thick layer of fibrous connective tissue
o With a covering of mesothelial cells
o Internally, this tissue subdivides the testis into
lobules
Lobules contain seminiferous tubules
Inside the testis
o Seminiferous tubules
Filled with spermatogenic cells
Spherical structures
Made up of specialized seminiferous epithelium with 2 types of cells
Sertoli cells (somatic cells)
Male sex cells (undergoing spermatogenesis and spermiogenesis)
Spermatogenic cycle
o In mice it lasts a total of 48 days
o Successive cycles start every 12 days
o Total of 4 overlapping cycles at every site along
the tubule

STRUCTURES
-

Inactive Sertoli Cells

Seminiferous tubules
o Structures found in seminiferous t.
Small blood vesses filled with red-stained
erythrocytes
P. Spermatocytes
Clumps of cells with conspicuous nuclei
that contains blue-stained chromatin
Interstitial cells/Leydig cells
S. Spermatocytes
o Produce testosterone
Connective tissue cells with elongated
nuclei
Sertoli Cells
o Nurse/sustentacular cells
o Only somatic cell in seminiferous tubule
o Spaced at regular intervals against basement membrane and around seminiferous tubules

Near spermatogonia
Have distinct outlines and large,oval
With indented nuclei with compound nucleoli
Surrounded by spermatozoa
Functions as:
Form blood-testes barrier
Nutrients and other
substances do
Spermatogonia
not directly reach cells
Determines which substances reach the
germ cells
Leydig Cells
Spermatogonia are outside bloodtestes barrier
Spermatids
P. Spermatocytes
Produce antigen-binding proteins
Necessary for spermiogenesis
o Two types of Sertoli Cells
Active Sertoli Cells
Normal function of supporting
spermatozoa
Pyramidal shape with apex towards lumen and nucleus perpendicular to tubule
wall
Inactive (resting) Sertoli Cells
Polygonal in shape
Nucleus is parallel to wall of tubule
Spermatogenic Cells
o Arranged in a layer
BASEMENT
o Vary in number with the phase of spermatogenic
MEMBRANE
activity
Does not take place in simultaneously in
all tubules
o The spermatogenic cells are:
GONIAL LAYER
Spermatogonia
Primary spermatocytes
Secondary spermatocytes
Spermatids
LUMEN
Spermatozoa
Spermatogonia
o Most primitive of the male sex cells
o Some undergo meiosis; progenies are translocated as
they
mature
Translocation mediated by sertoli cells
o Circumferential zones of more advanced cells inside zones
of less
advanced cells
o Criteria to identify spermatogonia
Rests on basement membrane and interrupted at intervals by sertoli cells
Gonial layer rapid mitosis (thin layers)
Nuclei is small, compared to spermatids
o 3 types of spermatogonia ***Near basement membrane except big triangular cells (sertoli)
1. TYPE A Spermatogonia
a. Chromatin appears fine and evenly dispersed
b. Single prominent chromatin nucleolus
o
o
o
o
o

ACTIVE

c. Ger
m
cell
line

Type B S. Spermatogonia

(eccentric position where u see the chromatin nucleolus)


d. Undergoes mitotic division
Late spermatid
2. Type B Spermatogonia
a. Chromatin is coarse granules or flakes of heavily stained chromatin
b. Most common gonia
Type A Spermatogonia
c. Undergoes meiotic division to become sperm cells
Intermediate S. Spermatogonia
d. Last stage before they transform into primary spermatocytes
3. Intermediate Spermatogonia
a. Chromophilic chromatin in the form of delicate crusts or flakes attached
to membrane
b. Nucleoli in central position
- Primary Spermatocytes
o Large size; large nuclei than the spermatogonia
o Visible chromosomes; nuclear membrane are disintegrated already
o Phases that can be identified on this level
Interphase
Granular nucleus with pale nucleolus
Primary spermatocytes have migrated inward
o Formation of new gonial layer multiplication of Type A Gonia
o Griffen and MacMahon
Estimated time of duration for this stage to be 3% of the total
time of the spermatogenic cycle as determined by Oakberg
o Oakberg
Cycle is 212 hours
Interphase exists for 6 hours
Metaphase I
Most easily recognized stage
20 discrete tetrads are visible
Tetrads are highly condensed
Aggregated at center (tetrads)
Anaphase I
Occasionally seen in section
Short duration
Spindle is distinct; present as dyads
Late anaphase shows cleavage furrow
Telophase I

Dyads aggregate at poles after anaphase


Cytoplasm constricts to form 2 cells
Nuclear membranes become distinct and dyads uncoil to form second meiotic
interphase
Secondary Spermatocytes
o Spermatogenic cells resulting from 1st meiotic division of primary spermatocyte
o Smaller than primaries
o Contain granular chromatin, globules of chromophilic chromatin free in the nucleoplasm or
associated with the nuclear membrane
o Resemble spermatids
o Twice the diameter of spermatids and half as numerous
Spermatids
o Resulting from division II lie inward beyond spermatocyte layer in seminiferous tubules
o Haploid
o Small nuclei
o Pale staining properties and above primary spermatocytes
o Nuclei elongates in time
o DNA undergoes condensation
o Nucleus decreases in size due to elimination of nuclear sap
o DNA increases in staining intensity as DNA becomes more and more condensed
o Early Spermatids
Golgi apparatus (or the acrosomal rudiment) appears as a red granule near nucleus
Flagellum appears early
Midpiece is a pink-stained thickening along first half of length
o Lie close to sertoli cells during transformation to spermatozoa
Spermatozoa
o Mature spermatozoa appear in sections as deeply stained short structures
o Innermost layer spermatogenic tubule
o Heads of most mature spermatozoa are buried in
sertoli
cells
o Tails extend into lumen
o Fully mature spermatozoa becomes free of
nurse cells and pass into ducts
o Mature spermatozoa found in lumina of
testis tubules and ducts is made up of:
Head (circle shaped and
flattened)
Middle piece
Tail (long and flagellated)

Chick Testis