Dr.M.M.

Misro

Reproductive processesFinal
output
Sperm is the finished product of the intricate

physiological processes
It is unique product in the sense that it is a
haploid cell, has a typical structure and ability to
move.
Shapes differ (species specific) but the basic
structure remains the same with head, mid
piece and tail.

Sperm-An unique cell

Sperms carry a net negative charge-the

distribution is more towards tail.

There are antigenic sites and lectin binding
sites all along the plasma membrane.
The antigenic domains are formed as far as
round spermatid stage but are altered,
reshaped, remodeled during transit through
epididymis.

Sperm
nucleus
The chromatin in sperm nucleus is highly
conserved.
The chromatin packaging is a specific type
DNA-protein association is different than the
somatic cell.
Histones are replaced with protamines in the
DNA of sperms which facilitates compact
packaging.

DNA Packaging

Sperm nuclear proteins

Up to the stage of round spermatids histones

are associated with the DNA.

Replacement of histones with protamines is
completed at the stage of elongated
spermatids.
There are two types of intermediate proteins,
TP1 and TP2 which get associated before the
final replacement with protamine takes place.

Sperm DNA-protamine
association

Such a specialized association is basically aimed

to provide greater protection to the chromatin
content, compact packaging requiring less space
and probably supports quick condensation and
decondensation processes.
The process gets completely reversed following
fertilization after the sperm enters into the
oocyte cytoplasm.

DNA-protein association changes

prior to fertilization
Sperm- DNA-Protamine (S-S)
Egg cytoplasm with lot of GSH converts DNA-

Protamine (-SH)
Nucleoplasmin in the oocyte has high affinity
for Protamine (-SH).
Histones found plenty in ooplasm replace
protamine
Sperm pronucleus-DNA-histone

Capacitation
Changes that take place are:
Rearrangement of plasma membrane proteins
Changes in the lectin binding pattern
Expression of new antigenic sites
Removal inhibitory factors known as
decapacitation factors

Acrosome reaction
In order to fertilize the sperm must

undergo acrosome reaction.

This is initiated when the sperm
comes in contact with the oocyte.
ZP3 is the zona pllucida glycoprotein
which acts as a ligand and its binding
with the sperm plasma membrane
receptors triggers the chain reactions
leading to acrosome reaction.
Such binding is only possible with
capacitated Sperms.

Sperm and seminal

Sperms are released in a semi viscous
plasma

medium-seminal plasma which forms

>95 % of the ejaculate. Sperm volume is
Just < 1%.
It is mainly secretions from accessory
glands seminal vesicle and prostateandrogen dependant organs.
Marker for seminal vesicle functionFructose.
Marker for prostate function-Acid
phosphatase and citric acid

Epididymis
Mammalian sperms are highly differentiated by the

time they leave testis.

But they do not have progressive motility and
fertilizing ability which they acquire while passage
through epididymis: Caput, Corpus and cauda.
Sperm maturation takes place in cauda.
Marker for epididymal function- neutral glucosidase

Epididymis (contd.)
It has fluid secreting and absorbing

capacities-estrogen receptors play a

significant role.
There is lot of adsorption of proteins from
these secretions on sperm plasma
membrane-in different regions new proteins
are added while few are taken out.
As a result, some of the intrinsic proteins
change their location during sperm matrn.
In men epididymis matrn. of sperm has little
significance because of rapid transport of
sperms from the epididymis.

Epididymis (contd.)
There is also active glycosylation of proteins in the

epididymis.
This supports the increase in net negative charge
and lectin binding ability.
Further chromatin condensation taken place in the
epididymis because of the increase in disulphide
bonding in DNA.
Epididymal spermatozoa get easily capacitated
compared to eja. Sperms.

Testis
These are paired of encapsulated organs

consisting of seminiferous tubules

separated by interstitial mass of tissue.
In most mammals the testis migrate from
their original site and pass through the
abdominal wall into an invagination of the
peritoneum-called scrotum.
The process of migration is called
testicular descent.
If the process fails, testis stays back in
the peritoneum-cryptorchidism-affecting
testicular function.

Testis(contd.)
The testicular capsule-tunica albuginea-

undergoes contraction-important for

maintaining interstitial pressure and sperm
transport.
In man-contracts once in every 14 min.responds to other stimulating agents like
acetyl choline, epinephrine, nor-epinephrine

Testis (Interstitial space)

It is the space between seminiferous

tubules.
Contains supplies of blood, lymph and
nerves of testicular parenchyma.
Cells include Leydig, mast cells,
macrophages.
Leydig cell has LH/hCG receptors and
androgen receptors
Leydig-macrophage-peritubular myoid cells
have established paracrine interactions.
Leydig cells-androgen synthesis and
release.

Testis-pituitary
interaction

Gonadotropins-LH and FSH are secreted from the

anterior pitutary following stimulation by GnRH

LH stimulated androgen production in Leydig
cells-FSH has receptors on Sertoli cells and germ
cells- The hormones are essential for initiation and
maintenance of spermatogenesis.

Anterior Pituitary
Hormones
Pituitary secretes-ACTH,TSH, GH, PRL,

LH,FSH
In addition to GnRH, hypothalamus
secretes a polypepide -PACAP.
Normally the enzyme adenylate cyclase
production is essential for cyclic AMP
production responsible for hormone
action. This peptide regulates
production and release of cyclic AMP in
the target cell population in the
pituitary.

MALE REPRODUCTIVE
SYSTEM
HORMONAL REGULATION OF
MALE REPRODUCTIVE FUNCTION
HYPOTHALAMUS REGULATES
ACTIVITY OF ANTERIOR
PITUITARY (ADENOHYPOPHYSIS)
ADENOHYPOPHYSIS
SYNTHESIZES HORMONES (LH and
FSH) THAT MODULATE ACTIVITY
OF SERTOLI AND LEYDIG CELLS
Luteinizing Hormone (LH): stimulates
testosterone production by Leydig cells
Follicle Stimulating Hormone (FSH): stimulates
production of sperm in conjunction with
testosterone by regulating activity of Sertoli
cells
SERTOLI CELLS STIMULATED BY FSH AND
TESTOSTERONE RELEASE ANDROGEN BINDING
PROTEIN WHICH BINDS TESTOSTERONE; THEREBY
INCREASING TESTOSTERONE CONCENTRATION
WITHIN THE SEMINIFEROUS TUBULES AND

 TESTIS

MALE REPRODUCTIVE
SYSTEM

TUNICA ALBUGINEA
- thick connective tissue capsule
- connective tissue
septa divide testis into
250 lobules
- each lobule contains 14 seminiferous tubules
and interstitial
connective tissue
(1) SEMINIFEROUS
TUBULES
- produce sperm
INTERSTITIAL TISSUE
- contains Leydig cells
which produce
testosterone
(2) RECTUS TUBULES
(3) RETE TESTIS
(4) EFFERENT
DUCTULES
(5) EPIDIDYMIS

Testis (seminiferous
tubule)
Testis mostly comprised of seminiferous
tubules. In rat, the diameter is 200-250 m
and lumen 50 m.
Tubules are double ended convoluted loops.
In rat there are about 30 tubules
In man-300 lobulations, each lobule
containing 1-4 tubules.

MALE REPRODUCTIVE TUNICA

ALBUGINE
SYSTEM
A
TESTIS

EPIDIDYMIS

Me
d
con iastin
u
TES tainin m
TIS g RE
TE

LOBULES

MALE REPRODUCTIVE
SYSTEM
TESTIS
TUNICA
VAGINALIS
TUNICA
ALBUGINEA
SEMINIFEROUS
TUBULES
SEMINIFEROUS
EPITHELIUM
- complex stratified
epithelium containing two
basic cell populations:
(1) SPERMATOGENIC
CELLS
(2) SERTOLI CELLS

MALE REPRODUCTIVE
SYSTEM
TESTIS
SEMINIFEROUS
TUBULES
SEMINIFEROUS
EPITHELIUM
- complex stratified
epithelium containing two
basic cell populations:
(1) SPERMATOGENIC
CELLS
stem cells which regularly
replicate and differentiate
into mature sperm as they
migrate toward the lumen
(2) SERTOLI CELLS
nonreplicating physical
support cells
INTERSTITIAL
CONNECTIVE TISSUE
(1) LEYDIG CELLS
produce and release

MALE REPRODUCTIVE
SYSTEM
SPERMATOGENESIS
SPERMATOGO
NIA

1
SPERMATOCYTE

2
SPERMATOCYTE

SPERMATIDS

SPERMATIDS
2 SPERMATOCYTE
SERTOLI
1 SPERMATOCYTE
CELLS:
- columnar with adjoining
lateral processes
- extend from basal lamina to
lumen
- Sertoli-Sertoli junctions
divide seminiferous tubules
into basal and adluminal
compartments

SERTOLI
CELLS

SPERMATOGONIA

Intratesticular
testosterone
The concentration of intratesticular testosterone

is very high.
In rat it is about 600 times more than the
peripheral circulation. In man-60-70 times.
The purpose of maintaining such a high conc. is
not very clear.
Therefore regulating the availability of this is vital
for complete arrest of spermatogenesis

Sertoli cell
This is the nongerminal component of the

seminiferous epithelium.
Sertoli cell proliferation stops after a
particular age, in rat 15-18 day after birth.
Provides a highly specialized environment.
Junctional complexes form the blood testis
barrier.
The barrier prevents the body in
recognizing cell surface antigens (sperms)
Prevents free flow of proteins,
immunocomplexes etc.

Sertoli cell
It secretes a variety of proteins-ABP, inhibin,

growth factor, transferrin, retinol binding

protein etc.
Provides the platform for germ cell
development and maturation.
The cytoplasm is extended both towards the
lumen and basal lamina. It has the capacity
to absorb the worn out cells.
Has the receptors for FSH

Spermatogenesis
The sequence of cytological events that result in

the formation of sperms from the precursor cellstermed-spermatogenesis.

Is a stage and phase specific process.
Intratesticular testosterone is maintained at a very
higher level.
Alterations in the production and availability of
testosterone affects spermatogenesis.

Spermatogenesis
There are five specific phases:
1. Spermatogonial or replication
2.
3.
4.

5.

phase
Meiotic phase
Acrosomal phase
Nuclear condensation and
elongation phase
Cytoplasmic elongation & release
phase

MALE REPRODUCTIVE
SYSTEM
SPERMATOGENESIS
THREE PHASES:
(1) Spermatogonial Phase
(Mitosis)
(2) Spermatocyte Phase
(Meiosis)
(3) Spermatid Phase
(Spermiogenesis)
- acrosome formation; golgi
granules fuse to form
acrosome that contains
hydrolytic enzymes which will
enable the spermatozoa to
move through the investing
layers of the oocyte
- flagellum formation;
centrioles and associate
axoneme (arrangement of
microtubules in cilia)
- changes in size and shape of
nucleus; chromatin condenses
and shedding of residual body

Spermatogenesis
One cycle of spermatogenesis takes 74 days in

humans, 48 days in rats and 34 days in mice.

Various orderly steps in spermatogenesis are:
spermatogonia, spermatocytes, spermatid and
spermatozoa.
The reduction division of spermatogenesis occurs
during primary spermatocyte stage.

Spermatogenesis
In rat there is segmental arrangement of stages,

where as in humans there is helical arrangement of

stages.
In a cross-section of the tubule, in rat only one

stage can be resolved, but in man there will be a

mixture of stages.

Spermatogenesis
There are two different types of

spermatogonia; one type forms cells that

undergo meiotic division and the other
type divide by mitosis and maintain the
number.
Spermatogonia are located outside the
blood testis barrier-spermatocytes once
formed are taken inside-reduction
division and further development of cells
take place inside the barrier.
In rat it consists of 13 stages & 19
spermatid steps- in man 6 stages and 8
spermatid steps.

MALE REPRODUCTIVE
SYSTEM

SPERMIOGENESIS
The process of transformation from round
spermatids to sperms-spermiogenesis.
Mature sperm 60m long and acquire full
motility in epididymis
(1) HEAD
- nucleus and acrosome
(2) NECK

- centriole and connecting

piece
(3) TAIL
- middle piece
(axoneme, outer dense fibers,
mitochondial sheath)

- principal piece

(axoneme, outer dense fibers, fibrous

sheath)

- end piece
(axoneme)

MALE REPRODUCTIVE
SYSTEM
SPERMIOGENESIS

Spermiogenesis
Takes about 3 weeks in man with following steps

Condensation of the nuclear DNA

The Golgi apparatus condenses to a single acrosomal granule situated

at the anterior portion of the nucleus.

Centrioles develop into the axoneme and other fibrillar elements of the
tail.
The volume of cytoplasm is reduced, and the mitochondria become
situated round the anterior portion of the tail elements (mitochondrial
sheath).
The spermatid cell membrane migrates caudally and forms the
cytoplasmic droplet near the neck of the spermatozoon.
The fully formed spermatozoon is then released into the seminiferous
tubule.
Qttn. is done from germ cells present during various stages.

Gonadotropin independent
spermatogenesis
Animal knockout models-hypogonadal hpg mouse

(GnRH deficient) shows reduced testicular size, low

intra-testicular androgen-germ cell arrest at
pachytene spermatocyte stage.
Exogenous testosterone completes
spermatogenesis despite undetectable FSH

FSH receptor knockout mice

It displays reduced fertility, testicular size, sperm

count and motility but a degree of completed

spermatogenesis.
FSH-B knockout mice exhibit normal fertility with
normal spermatogenesis but with 60% reduction in
Sertoli cell number and a marked decline in germ
cell number
It indicates, spermatogenesis, albeit at a reduced
level, is still possible in the absence of FSH action

LH Receptor knockout model

These mice are infertile, reduced testicular

size and hypoplastic accessary sex organs.

Germ cell development occurs but arrested
at the round spermatid stage.
Administration of Testosterone results in
resumption of spermatogenesis because of
AR activation but sperm number is low and
animals remain subfertile.
Similar results are obtained with LH
receptor knock out mice-spermatogenic
arrest at rST stage, low intra-testicular
testosterone levels.

AR knockout models
Complete AR knockout mice are infertile with

feminized phenotype, small cryptorchid testis,

spermatogenic arrest at pachytene
spermatocytes.
Some degree of germ cell development is
possible even in the absence of androgen action.
It is not clear whether second meiotic division can
always be completed or not

Gonadotropin independent
spermatogenesis in humans?
Patients with LH- subunit mutations

showed ongoing but severely impaired

spermatogenesis (Valdes-Socin et al,
2004).
But patient with FSH subunit mutation
displayed azoospermia.
Men with mutation of FSH receptor show
highly variable sperm densities (Phillip et
al, 1998)
Interesting observation comes from an
hypophy- sectomized men with activating
mutation in FSH receptor in which
spermatogenesis &fertility was
maintained despite absence of LH & FSH
(Gromoll et al, 1996)

Conclusion

The fine intricacies in the molecular steps

germ cell development under hormonal

regulation are already under intense
investigation in recent times.
Certainly, the concept of gonadotropin
independent spermatogenesis is worthy of
further consideration, particularly in
regard to developing contraceptives.
This necessitates further input in research
and development which will help us to
know more of such regulatory processes at
the molecular level.