ZOO 115 Animal Histology Laboratory Exercises

Exercise 12
THE REPRODUCTIVE SYSTEM
Male Reproductive System
The male reproductive system consists of the testes and their associated excurrent
ducts to conduct sperm to the outside. Usually also included in this system are various male
accessory glands which produce non-sperm components of the ejaculate; and that portion of
the urethra which is used for the transport of semen. It also includes the intromittent organ,
the penis.
OBJECTIVES: After completing the exercise on the male reproductive system, you should be
able to:
Identify the testis, epididymis, efferent ducts, and vas deferens in light microscopic
sections.
Identify Sertoli cells, primary spermatocytes, and spermatids in light microscopic
sections, and explain the relationship between the developing spermatozoon and the
Sertoli cell.
Identify and describe the function of testicular interstitial cells.
Identify the prostate gland in light microscopic sections.
Sketch or photograph structures viewed. Locate and label specific parts mentioned in
bold face.
1. Testis, Human H&E
This is good for studying the general structure of the organ. Use the lowest magnification
available. Identify the following: tunica albuginea, seminiferous tubules, interstitial CT and
interstitial (Leydig) cells.
The tunica albuginea, is the capsule forming the outer border of the testis demarcated by
a thick band of dense collagenous connective tissue. The connective tissue septa extending from
it subdivide the cavity of the testis into smaller compartments called lobules. Each lobule
contains 1-4 convoluted seminiferous tubules (about 150-300 m in diameter, 30-80 cm long).
Each seminiferous tubule continues near the mediastinum into a straight tubule, a tubulus rectus.
The straight tubules continue into the rete testis, a labyrinthine system of cavities in the
mediastinum, which is a conical mass of connective tissue from the tunica albuginea. The
mediastinum testis and rete testis are not visible in all sections.
In the regions between the seminiferous tubules (i.e., outside the adjacent boundary
layers) run small amounts of CT, and another population of cells is located here, one not part of
the germ line: the interstitial cells (or Leydig cells for Franz von Leydig; a German anatomist).
They are cuboidal in shape and present in groups of 2-20 cells. The cytoplasm is strongly
acidophilic and finely granular. The nucleus is large, round and often located eccentric in the
cell. These small groups are cell which produce the male steroid hormone testosterone. Their
location gives them their name.
2. Testis, rat/ mammal in H&E or PTS
Locate the seminiferous tubule. The insides of the tubules are lined with seminiferous
epithelium, which consists of two general types of cells: spermatogenic cells and Sertoli cells.
ATES-CAMINO, Fritzie B. (AY2011-2012), BSES-CSM, UPMindanao

ZOO 115 Animal Histology Laboratory Exercises

Identify Sertoli cells with their large pale nuclei and prominent nucleoli. It's a "nurse" cell that
sustains the proper environment for spermatozoa to develop. The Sertoli cells sits on the
outermost portion of the tubule, right up against the boundary layer: Its nucleus will be slightly
separated from the tube border, pale-staining, with an oval to pyramidal shape, and one or two
prominent nucleoli. The long axis of the nucleus is oriented perpendicular to wall of the
tubule. Although these cells have a very extensive and branching cytoplasmic structure, little of
the Sertoli cell's true extent can be seen in the light microscope.
Study the spermatogenic population in different stages of development.
Spermatogonia are basally located (i.e., at the periphery of the tubule) and can be
identified by their densely stained, round nuclei. These are diploid cells, and they're in the basal
compartment created by the Sertoli cells.
Primary spermatocytes are the initial product of spermatogonial maturation; you will
see these as large cells with a round nucleus filled with clumped chromatin material. Many of
them should be seen in various stages of the meiotic cycle.
Although secondary spermatocytes are undoubtedly present, these are very short-lived
and you're unlikely to tell it from a primary spermatocyte. By the time cells have progressed to
the spermatocyte stage meiosis is complete; hence secondary spermatocytes are haploid and
located in the luminal compartment.
The secondary spermatocytes further transform themselves into spermatids. Spermatids
are well on the road to final morphological differentiation termed spermiogenesis. They're
located much closer to the lumen of the tube, resting in the branches of the Sertoli cell
"tree." Spermatids will be seen in different stages of maturation. Some will be chunky and
squarish-looking cells with nuclei beginning to get quite dense, as at left. Others more advanced
will have tails beginning to develop. You may also see stages in which the excess cytoplasm is
being cast off. A "tailed spermatid" is a fairly common sight. A spermatid completing its
spermiogenesis is called a spermatozoon (pl. spermatozoa), which is a mature sperm having a
well-developed tail, neck, and head and found in the luminal region of the tubule.
3. Epididymis, human, H&E
The pseudostratified columnar epithelium contains two distinct cell types: tall
principal cells covered with stereocilia and smaller, basal cells which rest on the basal lamina.
In the lumen of the tubules there are some mature spermatozoa as well as cellular debris. In the
stroma making up the wall of the tubule, note connective tissue, smooth muscles and blood
vessels. The epididymis receives input of sperm from the efferent ducts and it acts as a storage
and maturation facility for them.
4. Ductus deferens, human
The ductus (vas) deferens is a thick-walled tube consisting of three concentric layers:
mucosa, muscularis and adventitia. The lumen of the duct is relatively small. The mucosa is
composed of a pseudostratified columnar epithelium with stereocilia (similar to the
epididymis) and a thin lamina propria rich in elastic fibers, which generally cause the mucosa
to form longitudinal folds. The muscularis is very robust and consists of three layers of smooth
muscle. The inner and outer layers have a longitudinal orientation, the intermediate layer
circular. The adventitia is continuous with the CT of the spermatic cord.

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ZOO 115 Animal Histology Laboratory Exercises

5. Human Penis, H&E

The ductus deferens carries sperm to the urethra, and the sperm are discharged through
that portion of the urethra which passes through the penis. Note the three cylinders of erectile
tissue: the two corpora cavernosa and the single corpus spongiosum containing the penile
(spongy) urethra. The erectile tissue is organized into paired dorsal corpora cavernosa and one
ventral corpus spongiosum. The corpus spongiosum around the urethra is also "erectile" tissue,
and its job is to keep the urethra patent during ejaculation. The tunica albuginea, a very dense
connective tissue surrounds both corpora cavernosa. The dorsal portion of the outer connective
tissue contains numerous blood vessels and nerves.
6. Male Accessory Reproductive Glands
The accessory (or secondary) male sex glands consist of the seminal vesicles, the
prostrate and the bulbourethral glands. In histological sections, most of these look much alike. Of
these glands, by far the most important to clinicians is the prostate gland, the largest accessory
sex gland in men.
A. Prostate, human or Mammal, H&E
The urethra, the opening in the center, is the major landmark by which to recognize this
gland. Examine the H&E appearance of the prostate gland. The smooth muscle content of the
fibromuscular stroma that characterizes the prostate is apparent in H&E sections.
The secretory alveoli of the prostate are very irregularly shaped because of papillary
projections of the mucosa into the lumen of the gland. The epithelium is cuboidal or columnar.
Basal cells are again present, and the epithelium may look pseudostratified where they are found.
The secretory cells are slightly acidophilic and secretory granules may be visible in the
cytoplasm. The secretory ducts of the prostate are lined by a simple columnar epithelium, which
changes to a transitional epithelium near the openings of the ducts into the urethra. A
characteristic feature of the prostate is the appearance of corpora amylacea in the secretory
alveoli. They are rounded eosinophilic bodies. Their average diameter is about 0.25 mm (up to 2
mm). Corpora amylacea can be seen in the slide of human prostate taken from an older
individual, but these concretions are not a distinctive feature in younger prostate or in the section
of monkey prostate. The secretion of the prostate is the first fraction of the ejaculate.
B. Seminal Vesicles, H&E
This is a highly folded, tubular gland, which primarily secretes fructose (and other
components) into the seminal fluid. The pseudostratified columnar epithelium contains tall,
nonciliated columnar cells and short, round basal cells that rest on the basal lamina. These
cells are apparently stem cells. The tall columnar cells have an abundance of rough endoplasmic
reticulum and large secretory vacuoles in the apical cytoplasm. There is an abundance of smooth
muscle arranged in two layers deep to the epithelium.
FEMALE REPRODUCTIVE SYSTEM
The female reproductive tract includes an egg-producing unit, the ovary, and a long tube
through which the discharged ova must travel (and in which fertilization takes place), the uterine
tube or oviduct. This latter organ can be subdivided into different regions, all of which are
physically continuous. The uterine tube leads to the uterus, the organ peculiar to Eutherian (or
"true", or "placental") mammals. It's in the uterus that gestation of the fetus occurs.
ATES-CAMINO, Fritzie B. (AY2011-2012), BSES-CSM, UPMindanao

ZOO 115 Animal Histology Laboratory Exercises

Following the normal period of development the term fetus is expelled through the
cervix, which may be considered as a separate component of the system, and eventually through
the vagina, the last component of the tract. This section focuses on the internal female
reproductive organs: the ovaries, oviducts, uterus and vagina. We will also have a look at
the mammary gland, an accessory reproductive gland. The external female genitalia include the
labia minora and majora, clitoris and vestibule.
OBJECTIVES: After completing the exercise on the female reproductive system, you should be
able to:

Identify the different stages of mammalian follicular development and degeneration in

microscopic sections.
Identify the regions of the mammalian ovary.
Identify the different portions of the uterine tube, based on their histologic appearance.
Identify the different regions of the female reproductive tract.

1. Ovary, Human, H&E

This section shows well the general features of the ovary. Identify the surface
(germinal) epithelium, tunica albuginea, hilum, cortex and medulla. The surface of the ovary is
covered by a single layer of cuboidal epithelium, also called germinal epithelium. It is
continuous with the peritoneal mesothelium. Fibrous connective tissue forms a thin capsule,
the tunica albuginea, immediately beneath the epithelium. Like so many other organs the ovary
is divided into an outer cortex and an inner medulla. The cortex consists of a very cellular
connective tissue stroma in which the ovarian follicles are embedded. The medulla is composed
of loose connective tissue, which contains blood vessels and nerves. Compare the stroma of the
medulla with that of the cortex. The cortical stroma consists of whorls of densely packed
fibroblastlike cells. The medullary stroma is more like loose fibroelastic connective tissue with
some smooth muscles cells. Follow the branches of the ovarian artery as they extend from the
hilum through the medulla and into the cortex.
2. Ovary, Cat or Human, H&E
The individual follicles, each of which contains an egg which can potentially be ovulated,
may be seen scattered through the cortex in various stages. Look for examples of each of the
following types of ovarian follicles (see figure, below): primordial, unilaminar primary,
multilaminar primary, secondary, mature (Graafian). Identify the following follicular features: 1
oocyte, zona pellucida, granulosa cells, theca interna, theca externa.
Primordial follicles. The primordial follicle is the earliest stage, the one in which all
follicles are at birth, and the one in which most of them die. The primordial follicle is
characterized by a single layer of thin, squamous follicular epithelium around its periphery.
Primordial follicles will be located close up under the outer surface of the organ (i.e., right up
near the tunica albuginea) usually occurring in clusters. They're grouped because when the
oogonium divides and redivides, the daughter cells tend to remain in the same area. Within each
primordial follicle is a primary oocyte (still diploid, because it's arrested in the course of its
meiotic division, in first meiotic prophase) surrounded by a single layer of squamous follicular
epithelial cells. The nucleus of the oocyte is positioned eccentric in the cell. It appears very light
and contains a prominent nucleolus.
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ZOO 115 Animal Histology Laboratory Exercises

Primary Follicle. The primary follicle is the first morphological stage that marks the
onset of follicular maturation. It also has a single layer of epithelium, but it's cuboidal to
columnar in shape. Their cytoplasm may have a granular appearance, and they are for this
reason also called granulosa cells. The continued proliferation of these cells will result in the
formation of a stratified epithelium (with a distinct basement membrane) surrounding the oocyte.
The zona pellucida (glycoproteins between interdigitating processes of oocyte and granulosa
cells) becomes visible. Parenchymal cells of the ovary surrounding the growing follicle become
organised in concentric sheaths, the theca folliculi. The oocyte becomes metabolically active,
and begins to synthesize new cytoplasm, causing it to enlarge compared to the oocyte in
primordial follicles, but it is still a primary oocyte.
Secondary Follicle. In this next stage, the follicular epithelium, called the granulosa
cells, becomes distinctly stratified, and the oocyte comes to develop a thick, translucent
covering, the zona pellucida (ZP). The ZP is an amorphous, non-cellular glycoprotein coat
surrounding the oocyte. which actually is a product of the surrounding follicular epithelium. It is
located between the oocyte and the follicle and the follicular cells. It serves as a sort of basal
lamina to the first layer of these cells, pressed up against the egg. Follicles demonstrating a
stratified follicular epithelium, and enclosing oocytes with a distinct ZP are secondary
follicles. Even so, the oocyte inside is still a primary oocyte. Two regions of cells can be
identified in the layer of granulosa cells surrounding the oocyte: the corona radiata, and the
cumulus oophorus. The corona radiata is the first layer of follicular (granulosa) cells outside the
zona pellucida. The cumulus oophorus is a thickened mound of granulosa cells that surround
the oocyte and projects into the antrum. Small fluid-filled spaces become visible between the
granulosa cells as the follicle reaches a diameter of about 400 m. These spaces enlarge and fuse
to form the follicular antrum, which is the defining feature of the secondary follicle. The
oocyte is now located eccentric in the follicle in the cumulus oophorus, where it is surrounded
by granulosa cells. Previously undifferentiated stromal cells, the theca folliculi now differentiates
with the continued growth of the follicle into two distinct layers around the developing follicle:
the theca interna and the theca externa. Cells in the theca interna are large, rounded and
epithelial-like. Vascularization of the theca interna improves, and the spindle-shaped or
polyhedral cells in this layer start to produce oestrogens. The theca externa retains the
characteristics of a highly cellular connective tissue with smooth muscle cells. The oocyte of the
secondary follicle reaches a diameter of about 125 m. The follicle itself reaches a diameter of
about 10-15 mm.
Mature or Graafian Follicle. The mature follicle, sometimes called the pre-ovulatory
follicle, has all of the components of the secondary-vesicular follicle but is much larger and
contains one single large antrum of follicular fluid. These follicles are very large and usually
extend from the deepest parts of the cortex and protrude from the surface of the ovary. In some
species just before ovulation, the primary oocyte in the mature follicle completes meiosis I
producing a secondary oocyte and a polar body.
3. Oviduct (Fallopian tube or Uterine Tubes), mammal or human H&E
The oviduct functions as a conduit for the oocyte, from the ovaries to the uterus.
Histologically,the oviduct consists of a mucosa and a muscularis. External to the mucosa and
muscularis is the serosa (visceral peritoneum).The mucosa of the uterine tubes is an
intermittently ciliated columnar epithelium that contains two types of cells: a ciliated cell and
a non-ciliated, secretory cell. The lamina propria consists of a typical loose areolar connective
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ZOO 115 Animal Histology Laboratory Exercises

tissue without glands, and it blends with the underlying, thin submucosa. There is no muscularis
mucosa layer in the entire female reproductive tract. The muscularis consists of an inner
circular and an outer longitudinal layer of smooth muscle. The serosa is typical containing
many blood vessels in a distinct vascular layer.
4. Uterus, H&E
The uterus is divided into body (upper two-thirds) and cervix. The wall is divided into
endometrium composed of the mucosal and submucosal layer, the myometrium composed of
fibromuscular layer, and the perimetrium, the serosal layer. The endometrium (mucosa of the
uterus) consists of simple columnar epithelium and lamina propria. The lamina propria is very
deep and made up of an extensive CT. A consistent feature of the lamina propria is the presence
of deep uterine glands. These glands provide nourishment for the early stages of embryonic
growth, before the placenta is established. Note the thick myometrium composed of
interconnecting bundles of smooth muscle arranged in four ill-defined layers. The
myometrium is the largest single piece of smooth muscle in the body, and its size and extent
can't really be appreciated in the microscope. The peritoneal surface of the uterus is covered by a
serosa the perimetrium, which probably may be lost on your slide.
5. Vagina, H&E
The vagina is a fibromuscular tube with a wall consisting of three layers: the mucosa,
muscularis and adventitia of the vagina. The mucosa, which may be thrown into folds, is lined
with stratified squamous non-keratinizing epithelium the thickness of which may vary with
the reproductive cycle. The epithelial cells are rich in glycogen (thus the empty appearance of
these cells in routine histological preps), which serves as a metabolic substrate for the
commensal bacteria of the vagina. The epithelium is supported by a loose to moderately dense
fibroelastic connective tissue stroma with abundant venous and lymphatic vessels. A
mucocutaneous junction is present where the lining meets the external integument of the body.
In the muscularis, inner circular and outer longitudinal layers of smooth muscle are present.
The part of the adventitia bordering the muscularis is fairly dense and contains many elastic
fibres. Loose connective tissue with a prominent venous plexus forms the outer part of the
adventitia.
6. Mammary Gland, H&E
The description here corresponds basically to the appearance of the resting mammary
gland. The mammary glands are modified glands of the skin. They are compound branched
alveolar glands, which consist of 15-25 lobes separated by dense interlobar connective tissue
and fat. Each lobe contains an individual gland. The secretory units are alveoli, which are lined
by a cuboidal or columnar epithelium. The excretory duct of each lobe, also called lactiferous
duct, has its own opening on the nipple. The lactiferous duct has a two layered epithelium - basal
cells are cuboidal whereas the superficial cells are columnar.

ATES-CAMINO, Fritzie B. (AY2011-2012), BSES-CSM, UPMindanao

ZOO 115 Animal Histology Laboratory Exercises

ATES-CAMINO, Fritzie B. (AY2011-2012), BSES-CSM, UPMindanao