Male Reproductive Physiology

Topics of Discussion
 Hypothalamic-Pituitary-Gonadal Axis
 The Testis
 The Epididymis
 Vas Deferens
 Seminal Vesicle and Ejaculatory Ducts
 Spermatozoa

HYPOTHALAMIC-PITUITARY GONADAL AXIS

 The hypothalamic-pituitary-gonadal (HPG) axis plays a critical role during development and
adulthood in 4 physiological processes:
1. Phenotypic gender development during embryogenesis
2. Sexual maturation at puberty
3. Testis endocrine function—testosterone production
4. Testis exocrine function—sperm production

2 kinds of hormones classes mediate intercellular communication here: Peptide and Steroid

Peptide hormones Steroid hormones

 Small, secretory proteins
 Act through cell surface receptors
 Hormone signals are transduced by second-
messenger pathways
 Ultimately, most peptide hormones induce
phosphorylation of proteins that alter cell
function.
 Example: luteinizing hormone (LH) and follicle-
stimulating hormone (FSH)
 Derived from cholesterol
 Not stored in secretory granules
 Secretion rates directly reflect rates of
production
 In plasma  usually bound to carrier proteins
 Because they are lipophilic  cell membrane–
permeable
 After binding to intracellular receptors 
translocated to nuclear DNA recognition sites
 regulate transcription of target genes
 Example: Testosterone and Estradiol

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 Hormonal signalling within HPG axis is governed by a free-running pulse generator within
hypothalamus.
 The specific amplitude and frequency with which hormone secretions occur within the reproductive
axis determine downstream organ responsiveness.
 Feedback control is the principal mechanism for hormonal regulation. With feedback control, a
hormone can regulate the synthesis and action of itself or of another hormone.
 In HPG axis, negative feedback is primarily responsible for maintaining homeostasis.

Hypothalamus
 Integrative center of HPG axis.
 Pulse generator for cyclical secretion of pituitary hormones.
 Receives neuronal input from Pons, Olfactory cortex, Retina, Thalamus, Amygdala, and many other
areas.
 Anatomically linked to pituitary by both a portal vascular system and neuronal pathways. By
avoiding the systemic circulation, the portal vascular system allows direct delivery of hypothalamic
hormones to anterior pituitary.
 M Imp hypothalamic hormone for reproduction = GnRH/LHRH
GnRH
 10–amino acid peptide.
 Secreted from neuronal cell bodies in preoptic and arcuate nuclei.
 T1/2 ~ 5 to 7 minutes. Almost entirely removed on 1st pass through pituitary (either by receptor
internalization or enzymatic degradation).
 Secretion from input from effects of stress, exercise and diet from higher brain centers,
gonadotropins from pituitary, and circulating gonadal hormones.
 GnRH stimulates the production and release of FSH and LH by a calcium flux–dependent mechanism.
 Sensitivity of the pituitary gonadotrophs for GnRH varies with age and hormonal status.

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 Substances known to regulate GnRH secretion

GnRH MODULATOR TYPE OF FEEDBACK EXAMPLES
Opioids Negative β-Endorphin
Peptide hormones Negative FSH, LH
Sex steroids Negative Testosterone
Catecholamines Variable Dopamine
Prostaglandins Positive PGE2
Insulin Positive Insulin
Kisspeptins Positive Kisspeptin (puberty)
Leptins Positive Leptin
He FELT Low (FSH, Endorphin, LH, T)
His Cat was Confused (Catecholamine)
He positively LIKEs to reproduce (Leptin, Insulin, Kisspeptin, E2 prostaglandin)
Kallman’s syndrome
 Congenital hypogonadotropic hypogonadism
 GnRH precursor neurons fail to migrate normally
 Subsequent absence of hypothalamic secretion of GnRH
 Affected individuals  delayed puberty or infertility (No T production)
GnRH output exhibits several types of rhythmicity:
 Seasonal: on a time scale of months and peaking in the spring
 Circadian: resulting in higher testosterone levels during the early morning hours
 Pulsatile: with GnRH peaks occurring every 90 to 120 minutes on average.
 Importance of a pulsatile secretory pattern is demonstrated by ability of exogenous GnRH agonists
(Leuprolide acetate) to # testicular T production (Continuous pituitary exposure to GnRH = negative
feedback)

Anterior Pituitary/Adenohypophysis
 Regulated by blood-borne factors (In contrast, Posterior lobe/neurohypophysis, is driven by neural
stimuli).
 Is the site of action of GnRH

LH and FSH
 Regulate testis function
 Glycoproteins: Composed of two polypeptide chains and one oligosaccharide.
 Polypeptide chain subunits:
1. α and β (coded by sep genes). Both subunits are required for endocrine activity.
2. α subunit of each hormone is identical & is similar to that of all other pituitary hormones.
3. Biologic and immunologic activities are conferred by unique β subunit.
 Sugars moiety = oligosaccharide with sialic acid residues
1. Differ in content between FSH and LH
2. Likely account for differences in plasma clearance of these hormones.
 FSH: Binds to Sertoli cells and spermatogonial membranes secretes ABP (Androgen Binding
Protein), Ceruloplasmin, Transferrin, Lactate, Clusterin, GF, PGs, Plasminogen activators. These
factors stimulate Seminiferous Tubule growth and stimulate spermatogenesis at puberty.

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LH FSH
Secretory Pulses 8-16/day Every 1.5 hr
Amplitude variation 1 – 3 times Max 25 % variation
of pulses
T1/2 Short Longer
Feedback T, E give negative feedback Activin, Inhibin
Secretion Closely resembles GnRH release Mainly GnRH independent (bcoz Activin
and inhibin are the main effectors)
Response of FSH to GnRH is difficult to
measure bcoz
 Activin and Inhibin
 Smaller amplitude of variation
Site of action Gonads Gonads
Action  Stimulate adenylate cyclase   Stimulate adenylate cyclase 
increase cAMP increase cAMP
 Stimulate T production  Bind to Sertoli cells and
(increased cholesterol  spermatogonial membranes 
pregnenolone T) stimulate Sem. Tubule growth
 Imp for initiation of spermatogenesis
at puberty
 Maintain normal spermatogenesis in
adults

Prolactin (Remember – Produces Lactin = Prolactin)

 Can also affect the HPG axis and fertility.
 Large, globular protein of 199 amino acids (23 kD).
 Responsible for milk synthesis in pregnancy and lactation.
 No human mutations have been found in either the gene or its receptor
 Normal role in men is less clear, but it may increase concentration of LH receptors on Leydig cells
and sustain normal, high intratesticular T levels.
 It may also potentiate the effects of androgens on the growth and secretions of the male accessory
sex glands.
 Normal Prolactin levels may be important to maintain libido.
 Although low Prolactin levels are not necessarily pathologic, hyperprolactinemia abolishes
gonadotropin pulsatility by interfering with episodic GnRH release
Additional Hormones
 ACTH, GH, and TSH can also have significant effects on male reproductive function.

TESTIS
 Interstitial compartment (Leydig cells) is responsible for steroidogenesis.
 Seminiferous tubules produce spermatozoa.

Testosterone
 Normal testosterone production in men is approximately 5 g/day.
 Secretion occurs in a irregular, pulsatile manner (nyctehemeral).
 Produced  secreted  Target tissue.

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 Metabolized into two major active metabolites in target tissue

1. DHT from the action of 5α-reductase
2. Estradiol through action of Aromatase
 DHT is a much more potent androgen than T.
 In most peripheral tissues, T reduction to DHT is required for androgen action
 In testis and skeletal muscle, conversion to DHT is not essential.
Protein hormones Inhibin and Activin
Inhibin
 32-kD protein made by Sertoli cells.
 Inhibits FSH release from pituitary (negative feedback at pituitary/hypothalamus).
 Within the testis, inhibin production is stimulated by FSH.
Activin
 Close structural homology to transforming growth factor–β
 Stimulatory effect on FSH secretion.
 Activin receptors are found in a host of extragonadal tissues, suggesting that this hormone may have
growth factor or regulatory roles in the body.
Negative feedback suppression of GnRH release
 Occurs through ARs in hypothalamus and pituitary
 Both testosterone(major) and estrogen(minor contribution) participate
 Testosterone feedback - Mainly at hypothalamus; T is primary regulator of LH secretion.
 Estrogen feedback - To pituitary; Estradiol (& inhibin) = predominant regulator of FSH secretion.

Development of HPG Axis

 Sex is genetically determined in humans
 Critical gene for sex determination = SRY (sex-determining region Y) on short arm of Y chromosome.
 SRY gene product = protein with a high mobility group box (HMG) sequence, a highly conserved
DNA-binding motif that kinks DNA. This DNA bending effect alters gene expression, leading to testis
formation and subsequently to the male phenotype.
 However, the SRY gene does not act in isolation.
 DAX1(nuclear hormone receptor gene) can alter SRY activity during development by suppressing
genes downstream to SRY (which normally induce testis differentiation)
 WNT4: largely confined to adult ovary, may also serve as an “antitestis” gene.
 The discovery of these genes has significantly altered theories of sex determination. In the past, the
female genotype was considered the “default,” SRY-negative, developmental pathway. It is now
clear that genes such as WNT4 and DAX1 can proactively induce female gonadal development, even
in presence of SRY
 Once gonadal sex is determined, Leydig cells make testosterone that induces development of
internal genitalia.
 Leydig cells also synthesize insulin-like-3 to promote transabdominal testis migration into scrotum
 DHT masculinizes genital anlage to form external genitalia
 Sertoli cells in developing testis synthesize müllerian-inhibiting substance (MIS, AMH) that prevents
the müllerian duct from developing into uterus and fallopian tubes and keeps the early germ cells
quiescent in testis
 Deficiencies in these developmental pathways generally result in either birth defects or intersex
disorders

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Early differentiation pathway of male Internal and external genitalia development

 The hormonal feedback relationships within the HPG axis also become established during gestation
 Expression of Kisspeptin protein is partly responsible for activating GnRH neurons and triggering
GnRH release.
 In addition, SF-1, an orphan nuclear receptor, is secreted by developing Sertoli cells and contributes
to HPG axis development.
 After the withdrawal of placental steroids at birth, there is a period of high gonadotropin secretion
in the neonate.
 Subsequently, as axis sensitivity to gonadotropins increases, FSH and LH secretions fall to the low
levels characteristic of childhood.
 Puberty begins with GnRH pulsing, leading gonadotropins to increase to adult levels and,
subsequently, to increase sex hormones.
 Hypothalamic capacity to generate GnRH pulses arises at puberty (~ 12th yr)
 Puberty begins at critical growth, weight, and nutritional rates for boys and girls, and is likely
initiated by Kisspeptin, melatonin, and Leptin.
Aging and HPG Axis (Male Menopause/Male Climacteric/Andropause/Partial Androgen Deficiency in
Aging Male (PADAM))
Decline in T
 Progressive decline in T and sperm production occurs with age.
 Men in 7th decade have mean T levels 35% lower than young men.
 T production is reduced because of
1. Decreased number of Leydig cells and
2. Increase in testosterone binding proteins
 Correspondingly, FSH levels also increase (mean = 3x higher in older men).
 Diurnal variation of T secretion is also lost.
Decrease in sperm production
 Is due to decrease in germ cell proliferation
 Changes in Seminiferous epithelium with age = decrease in ST volume and length.
Changes in HPG Axis response
 There is also evidence for a blunted HPG feedback response to Low T (despite generally high levels
of gonadotropins) and GnRH stimulation
 Finally, normal pulsatile GnRH release is replaced by irregular pulses that are less effective in
stimulating Gn release.

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TESTES
Anatomy
Gross
 White, ovoid organ
 Normal volume =15 to 25 mL
 Length = 4.5 to 5.1 cm
 Tunica albuginea has smooth muscle cells that
course through collagenous tissue.
 These smooth muscle cells
1. May impart contractile capability to
capsule
2. May affect blood flow into testis
3. Promote the flow of seminiferous
tubule fluid out of testis

 The testis parenchyma is divided into compartments separated by septa.

 Each septum divides seminiferous tubules into lobes that each contain a centrifugal artery.
 Seminiferous tubules harbour developing germ cells and interstitial tissue.
 Interstitial tissue is composed of Leydig cells, mast cells, macrophages, nerves, and blood and lymph
vessels.
 In humans, interstitial tissue comprises 20% to 30% of total testicular volume
 Seminiferous tubules are long, highly coiled, and looped.
 Both ends usually terminate in the rete testis.
 ~ 600 to 1200 tubules
 Estimated combined length ~ 250 meters
 The “hub” of the testis, also termed the rete testis, coalesces to form 6 to 12 ductuli efferentes that
carry testicular fluid and spermatozoa into the caput epididymis
 Relationship of interstitial tissue to seminiferous tubules
Arterial supply
 Arterial supply of testis and epididymis is derived from three sources:
1. External spermatic (or cremasteric) artery
2. Internal spermatic artery
3. Deferential (vasal) artery
 Internal spermatic artery arises from abdominal aorta and is intimately associated with pampiniform
plexus of veins.
 Vascular arrangement within pampiniform plexus, with counter-flowing artery and veins, facilitates
the exchange of heat and small molecules. T passively diffuses from veins to artery in a
concentration-limited manner. The countercurrent heat exchange supplies arterial blood to testis
that is 2° - 4° C lower than rectal temperature in normal men. Loss of temperature differential is a/w
testicular dysfunction in varicocele and cryptorchidism.
 Inferior to scrotal pampiniform plexus and near the mediastinal testis, spermatic artery is highly
coiled and branches before entering the testis.
 Extensive interconnections, especially between the internal spermatic and deferential arteries, allow
maintenance of testis viability even after division of the internal spermatic artery.

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 A single artery is observed in 50%, 2 arteries in 30% and 3 arteries in 20% of cases. From
angiographic studies, a single artery enters the testis in 56% of cases; two branches enter in 31% of
cases and three or more branches in 13% of testes.
 In men with a single testicular artery, its interruption can result in testicular atrophy.
 The testicular arteries penetrate tunica albuginea and then travel inferiorly along the posterior
surface of the testis within parenchyma.
 Branching arteries pass anteriorly over the testicular parenchyma.
 Major testicular artery branches also travel over the inferior pole of the testis, pass anteriorly and
branch out over the surface of the testis.
 The location of these vessels is clinically important, because they may be injured during Orchidopexy
or testis biopsy procedures.
 The midsection of the testis has relatively fewer vessels compared with superior or inferior areas
 Individual arteries to seminiferous tubules (centrifugal arteries) travel within the septa that contain
tubules.
 Centrifugal artery branches give rise to arterioles that supply individual intertubular and peritubular
capillaries.
 Intertubular capillaries: located within columns of interstitial tissue
 Peritubular capillaries: ladder-like capillaries running near seminiferous tubule
 Blood Flow ~ 9 mL/100 g of tissue/minute
Venous Drainage
 Veins do not run with corresponding intratesticular arteries
 Small parenchymal veins empty into either the veins on the testis surface, or into a group of veins
near the mediastinum testis that travels along the rete testis
 These two sets of veins join together with deferential veins to form pampiniform plexus as they
ascend into scrotum
 Pampiniform plexus veins are thin walled, which contributes to the passive diffusion of testosterone
and temperature exchange with spermatic artery.
Nerve Supply
 The testis has no known somatic innervation
 It receives autonomic innervation primarily from the intermesenteric nerves and renal plexus
 These nerves run along testicular artery
 Adrenergic innervation is restricted primarily to small blood vessels that supply Leydig cell clusters
(may regulate steroidogenesis)
Lymphatics
 Prominent lymphatics = observed within spermatic cord
 Interstitial space is drained by lymphatics, but not seminiferous tubules
 Obstruction results in dilatation of interstitium but not seminiferous tubules
 Lymphatic obstruction can also result in hydrocele, a known complication of Varicocelectomy and
Herniorrhaphy
 The sperm containing intratubular fluid that baths Sertoli cells flows from  seminiferous tubules
 rete testis  caput epididymis
 Isosmotic with plasma
 Mainly of seminiferous tubule origin
 Reabsorption in rete testis and efferent ductules is regulated by estrogens
 Tubular fluid composition is markedly different from blood plasma or lymphatics
 Substances are not freely diffusible into and out of tubules (“blood-testis barrier”)

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Testis Cytoarchitecture
INTERSTITIUM
 Interstitium contains blood vessels, lymphatics, fibroblasts, macrophages, mast cells, and Leydig
cells
Leydig Cells
 Responsible for testicular steroid production
 Differentiate from mesenchymal precursor cells by 7th week of gestation
 Activation of Leydig cell steroidogenesis correlates with onset of androgen-dependent
differentiation of male reproductive system.
 Leydig cells express steroidogenic enzymes before becoming responsive to LH
 They also differentiate from precursor cells under influence of
1. LH
2. placental-derived human chorionic gonadotropin (HCG)
3. Local paracrine factors such as IGF-1
 2 to 3 months after birth, a second wave of Leydig cell differentiation occurs in response to Gn
production from pituitary, briefly elevating testosterone levels in infants.
 Androgen produced during the early male neonate’s life is thought to hormonally imprint the
hypothalamus, liver, and prostate such that they respond appropriately to androgen stimulation
later in life.
 A single testis from a young adult contains approximately 700 million Leydig cells

Testosterone
 Synthesized from cholesterol
 Principal steroid produced by testis
 Numerous C18, C19, and C21 steroids are also produced
 The three main sources of cholesterol in Leydig cell are
1. Externally from blood-borne lipoprotein and internalization of cholesterol-lipoprotein
receptor complexes
2. De novo synthesis from acetate
3. Stored cholesterol esters in lipid droplets
 Maintenance of cholesterol stores is part of the normal resting function of the Leydig cell; LH
stimulation evokes cholesterol mobilization through cholesterol esterase activity.
 Pregnenolone is transported out of the mitochondrial membrane into the smooth endoplasmic
reticulum, where it is converted into testosterone. Testosterone diffuses across the cell membrane
and is trapped within the extracellular fluid and blood plasma by steroid-binding proteins.
 Cholesterol transport to inner membrane of mitochondrion is regulated by two transport proteins
1. Steroid acute regulatory protein (StAR) and
2. Peripheral benzodiazepine receptor (PBR)
 LH binding  cAMP  protein synthesis in Leydig cell newly-synthesized StAR contains a signal
sequence that enables it to be threaded through the outer mitochondrial membrane to facilitate
cholesterol transport.
 PBR forms a channel for cholesterol in mitochondrial membrane
 4 major enzymes in testosterone biosynthesis from pregnenolone are
1. Cholesterol side-chain cleavage enzyme
2. 3β-hydroxysteroid dehydrogenase
3. Cytochrome P450 17α-hydroxylase/C17-20-lyase
4. 17β-hydroxysteroid dehydrogenase

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Control of Testosterone Synthesis

 Involves both pituitary and nonpituitary factors
 Most important regulator is LH. Pituitary peptides other than LH (FSH and Prolactin) modify
response to LH
 Nonpituitary factors:
1. GnRH
2. Inhibin and activin
3. epidermal growth factor (EGF)
4. IGF-1
5. TGF-β
6. Prostaglandins
7. Adrenergic stimulation
8. Direct inhibition may also occur through estrogens and androgens

Testosterone Cycles
 T levels change dramatically during fetal, neonatal and adult life
 1st peak: in human foetus b/w 12-18 weeks of gestation
 2nd ~ 2 months of age
 3rd = 2nd or 3rd decade of life
 After this, there is a plateau, and then a slow decline with age
 Superimposed on this, there are annual and daily rhythms of testosterone production and irregular
daily fluctuations
 Peaks correspond temporally to following developmental events:
1. Differentiation and development of the fetal reproductive tract
2. Neonatal organization or “imprinting” of androgen-dependent target tissues
3. Masculinization of the male at puberty
4. Maintenance of growth and function of androgen-dependent organs in the adult

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Seminiferous Tubules
 Unique environment for gamete production
 Consist of germ cells and supporting cells
 Support cells include
1. Sertoli cells
2. Fibrocyte and myoid cells of basement membrane
 Germ cells include
1. A slowly dividing stem cell population
2. More rapidly proliferating spermatogonia and spermatocytes
3. Metamorphosing spermatids
Sertoli Cells
 Rest on tubular basement membrane and extend cytoplasmic ramifications into its lumen
 Ultrastructural features: irregularly shaped nuclei, prominent nucleoli, low mitotic index, and unique
tight junctional complexes between adjacent Sertoli cells
 These tight junctions are the strongest intercellular barriers in body
 They divide seminiferous tubule space into basal (basement membrane) and adluminal (lumen)
compartments
 This anatomic arrangement forms the basis for the “blood-testis barrier” and allows
spermatogenesis to occur in an immunologically privileged site
 Sertoli cells serve as “nurse” cells for spermatogenesis
 Undifferentiated spermatogonia are near basement membrane
 More advanced spermatocytes and spermatids  near the luminal surface.
A: Spermatogonia and early spermatocytes share
positions on the basal lamina and are enveloped
by adjacent Sertoli cells that join to form tight
junctional complexes (site of blood-testis barrier).
B: Sertoli cells form junctional complexes both
above and below leptotene-zygotene
spermatocytes as they translocate from the basal
to adluminal compartments.
C: The spermatocytes enter the adluminal
compartment when Sertoli tight junctions
dissociate.
D: The elongating spermatid is situated within a
narrow recess of Sertoli cell trunk.
E, As the spermatid elongates further, the
cell becomes lodged within the body of the
Sertoli cell. The advanced spermatid moves
toward the lumen of the epithelium in
preparation for spermiation. Only the sperm head
remains in intimate contact with the Sertoli cell.
Specialized cell-to-cell contacts: asterisks,
desmosome-gap junction complex; arrowheads,
ectoplasmic specializations; isolated arrows,
tubulobulbar complexes.

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 Thus the Sertoli cell is a polarized epithelium in which the base approximates the plasma
environment, and its apex harbours an environment unique to the seminiferous tubule
 Sertoli cells nurture germ cell development by:
1. providing a specialized adluminal microenvironment
2. supporting germ cells through gap junctions
3. allowing migration of developing germ cells within the tubule
 The tight junctions between Sertoli cells are constantly remodelled to allow “opening” and “closing”
necessary for germ cell interaction and migration
 Ligand-receptor complexes, such as c-kit and kit ligand, are likely involved in mediating
communication between germ and Sertoli cells
 Sertoli cells also participate in germ cell phagocytosis and produce and secrete fluid and important
effector molecules.
 Androgen-binding protein (ABP) is one of earliest described Sertoli cell secretory products. ABP is an
intracellular carrier of androgen within the Sertoli cell. By binding testosterone, ABP maintains high
levels of androgen (50-fold, as observed in serum) within the seminiferous tubules.
 Testosterone also plays an important role in the regulation of Sertoli cell function, including ABP
production.
 Inhibin is Sertoli cell–derived and plays an important regulatory role in the negative feedback loop of
FSH secretion. Inhibin B is emerging as an important endocrine marker of Sertoli cell function in the
male infertility evaluation.
 Sertoli cells maintain a germ cell microenvironment entirely distinct from that of plasma.
 Sertoli cells secrete other products including
1. Extracellular matrix components (lamin, collagen type IV, and collagen type I)
2. Proteins such as ceruloplasmin, transferrin, glycoprotein 2, plasminogen activator,
somatomedin like substances, T proteins, H-Y antigen, clusterin, cyclic proteins, growth
factors, and somatomedin
3. Steroids, such as DHT, testosterone, androstenediols, 17β-Estradiol, and numerous other
C21 steroids are also produced by Sertoli cells
Germ Cells
 Give rise to approximately 123 × 106 (range: 21 to 374 × 106) spermatozoa daily
 This equates to the production of about 1200 sperm per heartbeat.
 Within the seminiferous tubule, germ cells are arranged in a highly ordered sequence from the
basement membrane to the lumen.
 At least 13 recognizable germ cell types
 Each cell type = different step in spermatogenic process
 Proceeding from the least to the most differentiated
1. Dark type A spermatogonia (Ad)
2. Pale type A spermatogonia (Ap)
3. Type B spermatogonia (B)
4. Preleptotene (R)
5. Leptotene (L)
6. Zygotene (z)
7. Pachytene primary spermatocytes (p)
8. Secondary spermatocytes (II)
9. Sa, Sb, Sc, Sd1, and Sd2 spermatids
 Tight junctions maintain spermatogonia and early spermatocytes within the basal compartment and
all subsequent germ cells in the adluminal compartment.

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Ad = dark type A spermatogonium

Ap = pale type A spermatogonium
B = type B spermatogonium
II = secondary spermatocyte
L = leptotene spermatocyte
P = pachytene spermatocyte
R = resting/preleptotene 10 spermatocyte
Rb = residual body
Sa(a), Sb1(b1), sb2(b2), Sc(c), sd1(d1), Sd2(d2)
= spermatids
Z = zygotene spermatocyte
The table shows cells that make up the six
stages of the “cycle” of the seminiferous
epithelium (I to VI)
D1, diakenesis; ID and IID, first and second
maturation divisions of spermatocytes.

Peritubular Structure
 Seminiferous tubule is surrounded by several layers of peritubular tissue
 The outer adventitial layer consists of fibrocytes
 In the middle layer are myoid cells interspersed with connective tissue lamellae
 The inner layer consists of a collagen matrix
 Myoid cells are thought to have contractile function
 Myoid cells actively secrete extracellular matrix components fibronectin and collagen type I, and
produce the inner collagenous layer
 Myoid cells may also affect Sertoli cell function
 Skinner and coworkers (1988) isolated a paracrine factor produced by myoid cells, P-Mod-S
(peritubular modifies Sertoli), that profoundly affects Sertoli cell synthetic and differentiation
functions
 Human peritubular cells have also been shown to secrete testosterone, and exert regulatory Sertoli
cell secretory activity
Blood Testis Barrier
 More appropriately termed the “blood–seminiferous tubule barrier,” the barrier has two
components:
1. anatomic or mechanical element
2. functional elements
 The mechanical barrier is created, in part, by muscle-like myoid cells that surround seminiferous
tubules
 Regulation of molecular traffic also occurs at the level of capillary endothelial cells.
 most important component of this barrier is the synaptic tight junctions between Sertoli cells

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Why does the blood-testis barrier exist?

 Since it develops at spermarche, when spermatogenesis begins at puberty, it is likely important for
meiosis.
 In addition, it may immunologically isolate developing male gametes that are not recognized as
“self” by the adult male immune system
 the value of a blood-testis barrier is fully realized after puberty, because foreign “antigens” on
postmeiotic germ cells only exist after spermarche
 A testicular insult such as biopsy, torsion, or trauma will not induce antisperm antibodies if it occurs
before puberty
Spermatogenesis
 Total time to produce an ejaculated sperm ranges from 42 to 76 days
 Spermatogenesis involves
1. A proliferative phase as spermatogonia divide to replace their number (self-renewal) or
differentiate into daughter cells that become mature gametes
2. A meiotic phase when germ cells undergo a reduction division, resulting in haploid (half the
normal DNA complement) spermatids and
3. A spermiogenesis phase in which spermatids undergo a profound metamorphosis to
become mature spermatozoa.
 A cycle of spermatogenesis involves the division of primitive spermatogonial stem cells into
subsequent germ cells.
 Several cycles of spermatogenesis coexist within the germinal epithelium at any one time, and they
are described morphologically as stages.
 In addition, there is also a specific organization of spermatogenic cycles within the tubular space,
termed spermatogenic waves.
 The best evidence suggests that human spermatogenesis exists in a spiral or helical cellular
arrangement that ensures sperm production is a continuous and not a pulsatile process
Testis Stem Cell Migration, Renewal, and Proliferation
Testis Stem Cell Migration
 During early prenatal development, primordial germ cells migrate to the gonadal ridge and associate
with Sertoli cells to form primitive testicular cords
 These primitive germline stem cells are termed gonocytes after the gonad differentiates into a testis
by forming seminiferous cords
 They are called spermatogonia after migration to the periphery of the tubule
 these early migrating germ cells have properties very similar to embryonic stem cells and are likely
the source of adult seminoma germ cell tumors
Testis Stem Cell Renewal
 The growth factor-receptor kit ligand/c-kit receptor system and the niche factor glial cell line-
derived neurotrophic factor (GDNF) appear to be involved
 c-kit receptor is a marker of spermatogonial stem cells
 Spermatogonial stem cell renewal may be c-kit–independent
 Obtained from adult testis biopsies, the embryonic-like cells express distinct markers of pluripotency
(OCT-4, SOX-2, STELLAR, GDF-3), can form all three germ layers, maintain a normal karyotype, form
teratomas, and express appropriate levels of epigenetic markers and telomerase (Kossack et al,
2009). This finding suggests that, in the future, the testis may be a source of patient-specific,
embryonic stem cells for cell-based therapy.

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Testis Stem Cell Proliferation.

 In the human, pale type A (Ap) spermatogonia in the basal, stem cell niche of the seminiferous
tubule divide at 16-day intervals (Clermont, 1972) to form B spermatogonia.
 B spermatogonia are committed to become spermatocytes, but the cytoplasm between
spermatogonial daughter cells remains conjoined after mitosis, forming cytoplasmic bridges
between adjacent cells. These cytoplasmic bridges are observed between all classes of germ cells
throughout spermatogenesis (Ewing et al, 1980). These bridges could be important for synchronized
cellular proliferation and differentiation, and possibly for regulation of gene expression.
Meiosis
 The essential difference between mitotic and meiotic replication is that a single DNA duplication
step is followed by only one cell division in mitosis, but two cell divisions in meiosis (four daughter
cells).
 Consequently, daughter cells contain only half of the chromosome content of parent cells.
 Spermatogenesis begins with type B spermatogonia dividing mitotically to form primary
spermatocytes within the adluminal compartment
 Mature spermatocytes are the first germ cells to undergo meiosis
 A meiotic division is followed by a typical mitotic reduction division, resulting in daughter cells with a
haploid chromosome complement
 each daughter cell contains different genetic information
 The resultant cell is the Sa spermatid
 Chromosomal recombination, the defining feature of mammalian meiosis, ensures that haploid
gametes differ genetically from their adult precursors and is the real engine of genetic diversity and
evolution.
 Defects in the fidelity of recombination within human male germ cells can cause azoospermia and
male infertility
 In one study, 10% of nonobstructive azoospermic men had significant defects in recombination
compared to men with normal spermatogenesis
 In addition, among men with maturation arrest pattern on testis biopsy, faulty recombination was
observed in about half of cases
 Indeed, molecular evidence suggests that the correlation of faulty recombination and sperm
aneuploidy in azoospermic men is strong enough to explain the higher de novo rate of chromosomal
abnormalities in offspring conceived with in-vitro fertilization–intracytoplasmic sperm injection (IVF-
ICSI)
Spermiogenesis
 During spermiogenesis, round Sa spermatids mature into spermatozoa
 During this maturation sequence, cell division does not occur, but there are extensive changes to
the spermatid nucleus and cytoplasm.
 These include
o loss of cytoplasm
o migration of cytoplasmic organelles
o formation of the acrosome from the Golgi apparatus
o formation of the flagellum from the centriole
o nuclear compaction to about 10% of former size
o reorganization of mitochondria around the sperm midpiece
 The nucleus of the round spermatid changes from spherical to asymmetrical as chromatin
condenses.

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 Many cellular elements contribute to the reshaping process, including chromosome structure,
associated chromosomal proteins, the perinuclear cytoskeletal theca layer, the manchette of
nuclear microtubules, subacrosomal actin, and Sertoli cell interactions.
 With completion of spermatid elongation, the Sertoli cell cytoplasm retracts around the developing
sperm
 The mature sperm has remarkably little cytoplasm, and it is an elaborate, specialized cell produced
in massive quantity—up to 300 per gram of testis per second.
Sertoli Cell–Germ Cell Interaction
 A complex network of cell–cell interactions exists within the testis
o between Leydig cells and Sertoli cells
o between Leydig cells and peritubular cells
o between Sertoli and peritubular cells
o between Sertoli cells and germ cells
 Physical contact between these cells plays a role in propelling the germ cell toward the tubule lumen
and casting off of the residual cytoplasm from the spermatid.
 There are factors that can reversibly disrupt the blood-testis barrier, including transforming growth
factor–β3 (TGF-β3) and tumor necrosis factor–α (TNF-α)
 These substances act by reducing the levels of occludin and zonula occludens-1 (ZO-1) in the barrier
through a p38 mitogen-activated protein (MAP) kinase signalling pathway

Genetics
 Positional patterns of deletions (termed “microdeletions”) in the AZF region are used to subdivide
this region into AZFa, b, and c subregions
 Regional deletions of the Y chromosome, termed Yq microdeletions, occur in 6% to 8% of severely
oligospermic men and in up to 15% of azoospermic men
 Taken together, such deletions are the most commonly defined molecular cause of male infertility
 In contrast to partial and complete AZFc deletion patients, in which sperm is often found on semen
analysis or testis biopsy, the chance of finding ejaculated or testis sperm in men with complete AZFa
or AZFb deletions is highly unlikely
 Complete AZFa deletions are associated with germ cell aplasia or Sertoli cell–only histology
 Complete AZFb deletions are generally associated with maturation arrest at the primary
spermatocyte (early) or spermatid (late) stages
 AZFc deletions are associated with hypospermatogenesis or a Sertoli cell– only pattern with foci of
spermatogenesis.
 Sperm have been detected in ejaculates of men with presumed and confirmed partial AZFa and
AZFb deletions.
 Similarly, ejaculated sperm in men with AZFa + b, and AZFb + c deletions (presumably partial
deletions) has also been reported.
 AZFa−c deletions has been associated with azoospermia and no sperm on testis biopsy.

 More recently, it has become clear that the X chromosome may also be important for
spermatogenesis.
 Mutations in X-linked genes in male infertility patients, including the SOX3 gene (sex determining
region Y box 3) and the FATE gene.

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EPIDIDYMIS
Gross Architecture
 Comma-shaped organ
 Located along posterolateral surface of testis
 3 to 4 meters in length
 Tightly coiled and encapsulated within tunica vaginalis
 Extensions from sheath enter interductal spaces  form septa that divide the duct into
histologically characteristic regions
 Anatomically, these are classically divided into three regions:
1. Caput or head
2. Corpus or body
3. Cauda or tail
 Caput epididymis = 8 to 12 ductuli efferentes
from the testis.
 The lumen of ductuli efferentes is large and
irregular in shape near testis  becoming
narrow and oval near junction with ductus
epididymis.
 Distal to this junction, the duct diameter
increases slightly and, thereafter, remains
constant in the corpus epididymis.
 In the bulky cauda, the tubule diameter
enlarges substantially and acquires an irregular
shape.
 Progressing distally, the tubule gradually
assumes the characteristic appearance of the
vas deferens.
Blood Supply
 Caput and corpus - from a branch of
testicular artery. It subsequently divides
into superior and inferior epididymal
branches.
 Cauda by branches of deferential artery.
 Deferential and cremasteric arteries form
the collaterals to epididymis, when main
testicular artery is obstructed or ligated.
 Arterial branches enter along septa formed
from connective tissue sheath  coil
extensively  transform into straight
vessels of microvascular bed.
 Microvascularization density varies
significantly along the length of epididymis.
1. Proximal caput = densest
subepithelial capillary network.
2. More distal segments harbour less
dense vascularisation.
 Epididymal capillary network is under hormonal control

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Venous Drainage
 According to MacMillan (1954), venous drainage from corpus and cauda epididymis joins to form
vena marginalis epididymis of Haberer.
 These veins drain into pampiniform plexus through the vena marginalis testis/ cremasteric/
deferential veins.
Lymphatic drainage
 Occurs through two routes
 Lymph from the caput and corpus is removed through the same route as testis. These vessels course
beside internal spermatic vein  terminate in preaortic nodes.
 Lymph vessels from cauda join those draining vas deferens  terminate in external iliac nodes.
NS
 Pelvic plexus  inferior spermatic nerves
 Superior portion of hypogastric plexus  intermediate spermatic nerves
 Ductuli efferentes and proximal segments of epididymis are sparsely innervated by sympathetic
fibers
 Here, the fibers are observed in a peritubular plexus and are principally a/w blood vessels.
 Many more fibers are observed in the midcorpus.
 Density increases progressively distally coinciding with increase in number and proliferation of
smooth muscle cells.
 This of contractile cells and sympathetic nerves may explain the
1. Rhythmic peristaltic movements of ductuli efferentes and initial epididymal segments
2. Intermittent contractile activity of cauda and VD during emission.
 These physiologic contractions are critical to the movement of sperm through epididymis.
Cytoarchitecture
Epididymal Epithelium (Holstein -1969 and Vendrely -1981)
 It consists of two main cells types: principal cells and basal cells
Principal cells
 Vary in height along the length of epididymis due to length of stereocilia (microvilli, not cilia).
1. Tall stereocilia (120 μm) are generally found in proximal epididymis
2. Smaller or shorter stereocilia (50 μm) are observed in distal regions.
 Nuclei in PC are elongated, often possess large clefts and 1-2 nucleoli.
 PC carry out both absorptive and secretive processes.
 The cellular apices have numerous coated pits, micropinocytotic vesicles, multivesicular bodies,
irregularly shaped membranous vesicles and an extensive Golgi apparatus. These features vary along
the length of epididymis. Therefore, there is varying absorptive and secretory capacity along the
length of duct.
Basal Cells
 There are far fewer basal cells than PC lining the epididymal epithelium. Basal cells are dispersed
among principal cells.
 Tear-shaped basal cells rest on basal lamina and extend approximately 25 μm towards lumen, their
apices forming threads between adjacent PCs. They are thought to be derived from macrophages.
 Unlike PC, the morphology of basal cells remains relatively constant.
 They are thought to be the precursors of PC.

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Regional differences in epithelium along the length of the duct

 Within the epididymis proper, epithelium is pseudostratified and consists of PC and BC.
 More proximally, at the junction of the rete testis and ductuli efferentes, there is a distinct transition
from a low to a high cuboidal epithelium.
 Epithelium in ductuli efferentes appears uneven and consists of ciliated and nonciliated cells.
1. Ciliated cells - dispersed throughout epithelium; conduct sperm from efferent ducts into
epididymis.
2. Nonciliated cells with protruding apices - secretory in nature; predominate in proximal
ductuli efferentes.
3. Other nonciliated cells - have microvilli; s/o resorptive activity; predominate in distal ductuli
efferentes.
4. Nonciliated cells likely are involved in testis fluid reabsorption.
 Both nonciliated and ciliated cells are joined apically through junctional complexes. This suggests the
existence of a blood-epididymis barrier analogous to the blood-testis barrier. Although not as dense
as the blood-testis barrier, the blood-epididymis barrier extends from the caput to the cauda
epididymis and may play an important role in influencing the composition of fluid within different
segments of the epididymal lumen
Epididymal Contractile Tissue
 Outside the basal lamina of ductuli efferentes and epididymal tubule, there are various contractile
cells.
 In the ductuli efferentes, the contractile cells form a loose layer, two to four cells deep, around the
tubule. These cells contain myofilaments and are connected by numerous nexus-like junctions.
 In distal corpus, there are larger contractile cells with fewer nexus-like intracellular junctions that
resemble thin smooth muscle cells.
 In cauda, the thin contractile cells are replaced by thick smooth muscle cells that form three layers—
the outer two layers oriented longitudinally and the central layer circularly. This distal contractile
layer increases in thickness as it forms the vas deferens.
 The contractile tissue throughout the epididymis is likely involved in sperm transport.
Epididymal Function
 3 main functions
1. Sperm transport and storage
2. Sperm-fertilizing ability
3. Motility maturation
Sperm Transport
 ~ 2 to 12 days.
 ½ time is spent in caput-corpus epididymis, and rest ½ in cauda epididymis.
 Time spent is more likely related to daily testicular sperm production rather than a man’s age or the
frequency of ejaculation.
 In one study, sperm epididymal transit time averaged 2 days in men with a high daily rate of sperm
production (137 million per testis), compared with 6 days in men with low daily sperm production
(34 million per testis).
 Frequency of sex does not affect sperm transit time through caput and corpus but “recent
emissions” can reduce transit time through cauda by 68%.
 Sperms are immotile as they enter epididymis. They remain relatively immotile within caput.
Therefore, mechanisms other than sperm motility must exist to transport sperm through
epididymis.
 Other Mechanisms:

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1. Initially, sperm are carried into ductuli efferentes by rete testis fluid
2. Fluid flow is facilitated by fluid resorption by ductal epithelial cells mediated by Estrogen
Receptor.
3. Motile cilia and myoid cell contractions within ductuli efferentes also assist.
4. Within the epididymis proper, the principal mechanism responsible is spontaneous,
rhythmic contraction of contractile cells surrounding epididymal duct.
Sperm Storage
 After migrating through caput and corpus, sperms are retained in cauda for variable time,
depending on the frequency of sexual activity.
 In men (21 to 55 yr), ~ 155 to 209 million sperms/epididymis are present at any given point of time.
Approximately half are stored in caudal region.
 Spermatozoa in cauda (unlike testicular sperms) are
1. Capable of progressive motility and
2. Capable to fertilize eggs.
 Exact amount of time that sperm can remain fertile within the epididymis is unclear, but sperm can
remain viable for several weeks within cauda epididymis after VD ligation.
 But sperm fertility diminishes when maintained in the epididymis for prolonged time.
 Sperm aging occurs because of extended epididymal transit time and prolonged storage  reduced
fertility.
 Fate of unejaculated sperms - Exact fate is unknown.
 In animals, sperms are lost through
1. Spontaneous seminal discharge
2. Oral self-cleaning
3. Urine
4. Epididymal reabsorption.
 In humans
1. Phagocytosis of spermatozoa by macrophages (spermiophages) within the epididymal
lumen has been observed after ligation of VD.
2. However, whether this mechanism can remove large numbers of spermatozoa from the
epididymis of unvasectomized men is unclear.

Sperm Maturation – Motility , Fertility and Biochemical Changes

Sperm Motility
 As they pass through epididymis, sperm acquire increased capacity for motility.
 This is observed as both a change in the pattern of motility, and as an increase in the proportion of
sperm exhibiting “mature” motility patterns.
 Early epididymal Motility patterns
1. Most sperms are immotile or show only weak, twitching movement.
2. Occasional sperm shows “immature” tail movements characterized by “thrashing” beats in
wide arcs that result in little forward progression.
3. Proportion of sperm with immature motility pattern is higher in caput > corpus.
 Within corpus
1. There is an increase in fraction of sperm with a “mature” motility pattern, characterized by
high-frequency, low-amplitude beats that result in progressive motility.
 Within cauda
1. >50% of sperm have mature motility patterns
2. Remainder either immotile or showing the immature motility patterns

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Moore and colleagues (1983)

Region % Motility
Efferent duct 0
Immature pattern
Caput 3

Proximal Corpus 12

Distal Corpus 30
Mature pattern
Cauda Epididymis 60

Sperm Fertility
 Testicular sperm are incapable of fertilizing
eggs unless injected into them with
micromanipulation.
 The ability to fertilize eggs is acquired gradually
as sperm pass through distal epididymis.
 Sperm from proximal epididymis are able to
bind to zona-free eggs.
 Only sperm from cauda epididymis can both
bind and penetrate eggs.

Sperm Biochemical Changes

 Sperm undergo many biochemical changes in epididymis.
 Epididymal transit induces a net negative surface membrane charge  Sperm membrane sulfhydryl
groups oxidize to disulfide bonds  improving sperm structural rigidity necessary for progressive
motility and egg penetration.
 Other post-testicular modifications of sperm membranes which enhance sperm adherence to egg
zona pellucida include changes in
1. Sperm lectin-binding properties
2. Phospholipid and lipid content
3. Glycoprotein composition
4. Immunoreactivity and
5. Iodination characteristics.
 Metabolic changes during epididymal transit
1. Increased capacity for glycolysis
2. Changes in intracellular pH and calcium content
3. Modification of adenylate cyclase activity and
4. Alterations in cellular phospholipid and phospholipid-like fatty acid content.

Regulation of Epididymal Function

 Sperm changes within epididymis are likely influenced by fluids and secretions within epididymal
lumen.
 Biochemical composition of epididymal fluid not only differs from that of serum, but there are also
regional differences in the osmolarity, electrolyte content, and protein composition.

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 These differences are due to variations in

1. Vascularization
2. Blood-epididymis barrier activity and
3. Selective absorption and secretion of fluid constituents such as glycerylphosphorylcholine
(GPC), carnitine, and sialic acids along epididymal duct.
 Proteins within the epididymal fluid that are known to have physiologic effects on sperm
1. Forward motility protein
2. Sperm survival factor
3. Progressive motility sustaining factor
4. Sperm motility-inhibiting factor
5. Acidic epididymal glycoprotein and
6. EP2-EP3 proteins (induce sperm binding to zona pellucida)
 Thus variations in epididymal tubule fluid characteristics play an important role in sperm maturation
during epididymal transit. It is not surprising, then, that epididymis is a potentially important source
of sperm dysfunction and male infertility.
 Hormonal Regulation
1. Epididymal function is hormonally regulated.
2. T and DHT are found in very high concentrations within epididymis, and there are no
regional gradients in androgen levels.
3. Bilateral castration results in
 Loss of androgen-dependent epididymal proteins
 Loss in epididymal weight
 Changes in luminal histology
 Alterations in synthesis and secretion of GPC, carnitine and sialic acid.
 Ultimately, it loses the ability to sustain sperm motility, fertility maturation, and
sperm storage capacities.
 Most of these degenerative processes are reversed with androgen replacement.
4. Compared with other accessory sex glands, epididymis requires relatively higher levels of T
and DHT for maintenance of its structure and function.
5. Effects are mediated mainly through DHT and/or 5α-androstane-3α, 17β-diol (3α-diol).
6. This is corroborated by the fact that the enzymes 5α-reductase (TDHT) and 3α-
hydroxysteroid dehydrogenase (DHT3α-diol) are found in the human epididymis.
 Epididymal function is also influenced by temperature. Chronic exposure to elevated temperatures
(example varicocele/ cryptorchidism) loss of sperm storage and electrolyte transport functions.
 Abnormalities in epididymal myoid cell contractility may also influence epididymal function.
1. Partial surgical denervation of epididymis  abnormal accumulation of sperm within cauda
+ decrease in swimming speed.
2. This becomes important in infertility due to neuropathic causes such as SCI and DM.

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VAS DEFERENS
Gross Architecture
 Tubular organ derived from MND
 30 to 35 cm long. Begins at cauda epididymis and terminates in ED, medial to SV and posterior to
prostate.
 It is classically divided into 5 regions:
1. Sheathless epididymal segment contained within tunica vaginalis
2. Scrotal segment
3. Inguinal segment
4. Retroperitoneal or pelvic portion and
5. Ampulla
Histology
 Outer adventitial connective tissue sheet - containing blood vessels and small nerves
 Muscular coat
1. Inner longitudinal layer
2. Middle circular layer
3. Outer longitudinal layer
 Inner mucosal layer - with an epithelial lining
 Outer diameter varies from 1.5 to 3 μm
 Lumen of unobstructed VD varies from 200 to 700 mm in diameter
 BS - Deferential artery, a branch of the superior vesical artery
 Venous drainage - corresponds to arterial supply.
 NS - both sympathetic and parasympathetic
1. Cholinergic supply does not appear important for motor activity
2. There is a rich supply of sympathetic adrenergic nerves derived from hypogastric nerve
coursing via presacral nerve.
3. Adrenergic nerve fibers have been observed in all 3 layers of vas tunica muscularis (greatest
concentration in outer longitudinal layer)
4. VD also receives a short adrenergic nerve
5. VD also has an abundance of ligand-gated, purinergic receptors in its smooth muscle
membranes, suggesting sympathetic and purinergic cotransmission in sperm transport and
ejaculation.
6. Neurons containing other NTs, including neuropeptide Y, enkephalin, galanin, somatostatin,
VIP, and NO have also been identified - however, their role is unknown.
Changes in VD - 1 to 15 years post-vasectomy
 Marked reduction in density of muscular noradrenergic and subepithelial secretomotor nerves in
testicular compared to abdominal segments.
 These changes may influence subsequent sperm maturation and transport, and hence procedural
success after vasectomy reversal.
Cytoarchitecture
 Lined by pseudostratified epithelium
 Height of epithelium decreases along the length of VD from testis to SV.
 Longitudinal epithelial folds are simpler proximally near testis and become more complex distally.
 Pseudostratified lining is composed of basal cells and 3 types of tall, thin columnar cells.
 Columnar cells
1. Extend from epithelial base to lumen
2. Include principal cells, pencil cells and mitochondria-rich cells.

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3. All columnar cells exhibit stereocilia and irregular convoluted nuclei.

4. Principal cells are MC columnar cell type in proximal VD
5. Both pencil cells and mitochondria-rich cells increase in density distally.
 Thickness of total muscle layer gradually decreases along the length of VD.
Vas Deferens Function
 3 main functions
1. Sperm Transport
2. Storage
3. Absorption and Secretion
Sperm Transport
 Sperm transport through VD is influenced by several physiologic processes.
 First, VD exhibits spontaneous motility.
 2nd, it also has the capacity to respond when stretched
 3rd, fluid within VD can be propelled into urethra by strong peristaltic contractions elicited either by
electrical stimulation of hypogastric nerve or by adrenergic NTs.
 This suggests that immediately before emission, with sympathetic stimulation, rapid transport of
sperm from distal epididymis  VD  ED occurs.
 This rapid transport ability is there, because VD has greatest muscle to lumen ratio (≈10 : 1), of any
hollow viscus in the body.
Storage
 Sperm reserves in VD ~ 130 million/VD.
 Vasal sperm quality - is very similar to ejaculate; 71% motility and 91% viability.
 During sexual rest - epididymal sperm are transported through VD and leak into urethra in small
amounts at irregular intervals. This helps in getting rid excess, stored sperm.
 On sexual stimulation - sperm are transported through VD.
 After sexual stimulation - distal VD contracts with greater amplitude, frequency and duration than
the proximal VD  contents are propelled back towards the epididymis.
 With prolonged sexual rest - excess epididymal sperm are once again transported distally.
 Therefore, VD is important not only for sperm transport, but also for maintenance of epididymal
sperm reserves.
Absorption and Secretion
 VD has both absorptive and secretory functions.
 Principal cells
1. Typical cells that synthesize and secrete glycoproteins.
2. The stereocilia, apical blebbing, and primary and secondary lysosomes within principal cells
are also characteristic of cells involved in absorptive function.
 Epithelial cells
1. In the ampulla of VD
2. Show spermiophagy
Regulation of VD function
 Normal VD function is likely to be androgen dependent
 VD actively converts testosterone to DHT.
 Castration causes atrophy of vas cytoarchitecture.
 Testosterone treatment causes restoration of vas cytoarchitecture.
 Spontaneous and α- and β-adrenergic–stimulated contractions of VD are altered by castration.
 Thus, once thought to be a simple muscular conduit for sperm, VD is now viewed as a complex
reproductive organ.

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SEMINAL VESICLE AND EJACULATORY DUCTS

Gross and Cytoarchitecture
Seminal Vesicle
 In the adult, the seminal vesicles are paired, elongated, hollow viscous organs located posterior to
the prostate and bladder.
 Each SV is 5 to 7 cm long and up to 1.5 cm wide.
 Each SV consists of a tubule that is 15 cm long and highly coiled and convoluted.
 Tubule is composed of three layers: inner, middle and outer.
1. Inner lining: is a moist and folded mucous membrane
2. Middle layer is largely collagenous
3. Outer layer consists of circular and longitudinal muscles layers that constitute 80% of the
wall thickness.
 Mucosa:
1. Mainly nonciliated, pseudostratified columnar or cuboidal cells
2. Notable for many thin, complicated folds that produce numerous crypts.
 Excretory duct of SV opens into ampulla of VD as it enters prostate.
 BS:
1. Arises from Internal iliac and inferior vesicular artery through prostatovesicular branch.
2. PV artery can also arise from superior vesicular/pudendal artery.
3. PV artery has anterior and posterior branches that supply the respective surfaces of SV.
 LD : Internal iliac lymph nodes.
 NS :
1. Sympathetic nerves from superior lumbar and hypogastric nerves.
2. Parasympathetic innervation occurs through the pelvic plexus.
Ejaculatory Ducts
 Paired, collagenous, tubular structures that commence at junction of VD and SV
 Course through prostate
 Empty into prostatic urethra at verumontanum.
 Histologically, EDs are a continuation of SV, except that the outer circular muscle layer does not
extend into ducts.
 3 distinct anatomic regions:
1. Proximal, extraprostatic portion
2. Middle intraprostatic segment
3. Short distal segment incorporating the lateral aspect of verumontanum in urethra.
 Although ED contains an outer muscular layer in its extra- and intraprostatic segments, as the duct
courses distally, the outer muscular layer dissipates, and there is no valvelike, muscular “sphincter”
at the ejaculatory duct orifice, as was once thought.
 Instead, urinary reflux is prevented and ejaculatory continence is maintained by the acute angle of
duct insertion into the urethra.
 Inner epithelial layer of ED is also complex and folded, and consists of simple and pseudostratified
columnar cells.
 BS: Branches of inferior vesical artery
 NS: Pelvic plexus.

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Seminal Vesicle and Ejaculatory Duct–Unit Function

 Functionally similar to bladder and urethra.
1. SV = contractile, compliant, smooth muscular organ with dynamic properties like bladder.
2. ED serve as a urethra-like conduit.
This theory allows the classification of ejaculatory duct obstruction into two types of disorders,
analogous to BOO
(1) obstruction resulting from physical blockage of ED, similar to BOO
(2) “functional” obstruction of SV, similar to voiding dysfunction due to bladder myopathy.
In addition, this has implications for diagnosis of ejaculatory duct disorders because “static” anatomical
imaging, such as TRUS, may not be sufficient to differentiate between these disorders, and medications
and conditions (such as diabetes) might predispose the system to seminal vesicle dysfunction.
Seminal Vesicle Function
 Secrete significant proportion (80%) of seminal fluid, and these secretions are found in later
fractions of the ejaculate, after the sperm-rich epididymal, and prostatic secretions.
Role in capacitation:
 After ejaculation, sperm pass into and through female cervical mucus  uterus  oviduct 
fertilization.
 During residence in female reproductive tract, sperm must undergo capacitation prior to oocyte
fertilization.
 Capacitation occurs at different rates for individual sperm. During capacitation, the acrosome
reaction and development of hyperactivated motility occurs.
 It is not clear if prostatic or seminal vesicle secretions contribute to capacitation.
Role of SV Fluid
1. Exact physiologic role is not clear
2. May function as a plug or barrier that reduces the chances for sperm from a subsequent male to
fertilize the oocyte.
3. Before ejaculation, semen is a liquid. After all components mix with the seminal vesicle secretions, it
coagulates. The major component of the coagulum is semenogelin I, a 52-kD protein expressed
exclusively in SV. By coagulating semen, SV secretions may promote sperm motility, increase
stability of sperm chromatin, and suppress immune activity in female reproductive tract.
4. The best-elucidated function of human semen appears to be its ability to provide antioxidative
protection to sperm. Semen is rich in antioxidant enzymes, including glutathione peroxidase,
superoxide dismutase, and catalase. In addition, the antioxidant molecules taurine, hypotaurine,
and tyrosine are present in high concentrations.
5. Lipofuscin granules from dead epithelial cells give seminal vesicle secretions a yellow-white color.
6. SV secretions are alkaline and contain fructose, mucus, vitamin C, flavins, phosphorylcholine, and
prostaglandins.
7. High fructose levels provide nutrient energy for the sperm when studied in vitro.
8. Mixing of SV with prostatic secretions results in human semen having a mildly alkaline pH.
 Acidic ejaculate (pH <7.2) is associated with blockage or absence of seminal vesicles

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SPERMATOZOA
 A remarkably complex metabolic and genetic machine
 Approximately 60 μm in length
 Divided into: Head, neck, and tail
Head
 Oval
 About 4.5 μm long and 3 μm wide
 Contains a nucleus with highly compacted chromatin, and an acrosome, a membrane-bound
organelle that harbours enzymes required for penetration of the outer vestments of the egg before
fertilization
Neck
 Maintains the connection between the sperm head and tail
 It consists of connecting piece and proximal centriole
 Axonemal complex extends from proximal centriole through the sperm tail.
Tail
 Midpiece, principle piece, and end piece
 Midpiece
1. 7 to 8 μm long
2. Most proximal segment of the tail
3. Terminating in annulus
4. It contains the axoneme, which is the 9 + 2 microtubule arrangement, and surrounding
outer dense fibers
5. It also contains the mitochondrial sheath, which is helically arranged around the outer dense
fibers.
6. The outer dense fibers, rich in disulfide bonds, are not contractile proteins but are thought
to provide the sperm tail with the elastic rigidity necessary for progressive motility
 Principal piece
1. Similar in structure to Midpiece
2. Has several columns of outer dense fibers replaced by fibrous sheath.
3. Fibrous sheath consists of longitudinal columns and transverse ribs.
 Endpiece
1. The sperm terminates in end piece
2. Contains axonemal structures and the fibrous sheath.
 Except for the end-piece region, the sperm is enveloped by a highly specialized plasma membrane
that regulates the transmembrane movement of ions and other molecules
Sperm Mitochondria
 75 mitochondria surround the axoneme
 Semiautonomous organelles
 Contain enzymes required for oxidative metabolism and produce ATP
 Also cause apoptotic cell death through release of cytochrome c.
 5 respiratory chain complexes span width of inner membrane (Oxidative Phosphorylation)
1. Nicotinamide adenosine diphosphate (NADPH) dehydrogenase
2. Succinate dehydrogenase
3. Cytochrome bc1
4. Cytochrome c oxidase, and
5. ATP synthase complexes.

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 Contained within the matrix are citric acid cycle, fatty acid and amino acid oxidative enzymes, newly
made ATP, mitochondrial DNA (mtDNA), and ribosomes.
Plasma membrane covering the sperm-head harbours specialized proteins
 Participate in sperm–egg interaction
 Carbohydrate-binding proteins:
1. Interact with species-specific ZP3 protein in egg zona pellucida  sperm binding to zona 
induction of acrosome reaction
 PH30:
1. Modified during sperm migration through epididymis
2. Functions as a fusion protein b/w sperm and egg membranes
Axoneme:
 Physiologically, it is the true motor assembly
 requires 200 to 300 proteins for proper function.
 Contains enzymes and structural proteins for chemical transduction of ATP into motility.
 Microtubules are the best understood components. They are arranged in classic “9 + 2” pattern (9
outer doublets encircling an inner central doublet)
 The complex protein Dynein extends from one microtubule doublet to the adjacent doublet, and
forms both the inner and outer “arms” of the axoneme.
 Dynein
1. Large (2000 kDa), Mg+-stimulated, ATPase
2. Responsible for ATP-generated microtubule sliding  causes axonemal bending  flagellar
movement
 Heads
1. 2-3 globular, outer (heavy) chain heads (500 kDa) joined to a common stem.
2. The heads control movement along the microtubules.
 Arms
1. The inner (light) chain arms(14 to 120 kD)
2. primary effectors of movement
3. associated with the radial spokes of the dynein assembly.
4. Sperm with outer arm mutants have reduced motility, and those with inner arm mutants
have no motility.
5. Radial links or spokes connect a microtubule of each doublet to the central inner doublet
and consist of a complex of proteins.
6. The central inner doublet is surrounded by a ringlike helical sheath to which the radial links
from the outer doublets are attached.
 Tektins are proteins associated with the outer microtubular doublets
 Nexin links are proteins that connect the outer doublets to each other and maintain the cylindrical
axonemal shape.
 Human mitochondrial DNA (mtDNA)
1. is distinct from the sperm nuclear DNA
2. consists of a circular, histone-free chromosome of 16,569 bp of DNA arranged in a single
heavy and single light strand.
3. MtDNA encodes 13 respiratory-chain-complex subunit proteins, 2 mitochondrial rRNAs, and
22 tRNAs used for protein synthesis. These genes have no introns.
4. Expression of mtDNA genes are regulated by strand-specific, but not gene-specific,
promoters in response to activation of a transcription factor (mtTFA).
5. far more susceptible to mutations than nuclear DNA (estimated 40 to 100 times higher).

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6. Reasons for this may include the fact that mitochondria are near respiratory-chain
complexes and may be easily attacked by reactive oxygen species.
7. In addition, mtDNA is not coated with protective histones, and mitochondria have very
limited DNA repair mechanisms
8. The fact that mitochondria rapidly accumulate mutations suggests the necessity of
degrading all paternal mtDNA in the fertilized egg. This degradation is likely mediated by the
small proteolytic polypeptide ubiquitin that regulates proteolysis in many tissues

Ciliary dyskinesia
 “defective sperm structure”
 Although infertility is the rule with ciliary dyskinesias, ejaculated sperm can be motile and sperm
concentrations can be normal.
 With ICSI, clinical pregnancies and live births have been reported using affected sperm
 Because the inheritance is usually recessive, normal offspring are likely.
 Patients suspected of harbouring sperm structural defects generally exhibit severely compromised
sperm motility (<10%).
 Sperm electron microscopy can reveal ultrastructural or functional abnormalities of the sperm.

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Sperm structural abnormalities: Categorized by Chemes (2000)

Nonspecific Flagellar anomalies Dysplasia of fibrous sheath
MC Flagellar anomaly a/w severely low motility Systematic sperm abnormality
Usually associated with near-total or total
immotility.
Random, heterogeneous, microtubular alterations Homogenous fibrous sheath, axonemal, and
periaxonemal distortions.
No e/o familial occurrence Strong familial incidence
Sometimes due to correctable disorders such as A subset have classical ciliary dykinesias (formerly
varicocele, reactive oxygen species, and immotile cilia syndrome) = sperm immotility +
gonadotoxin exposure. respiratory disease + dextrocardia
 Sperm are ciliated cells that possess a “9 + 2” axonemal structure that allows motility.
 It is estimated that 200 to 300 genes regulate sperm motility.
 Sperm motility defects, termed ciliary dyskinesias, are common and can be either correctable
(nonspecific flagellar anomalies) or genetic (dysplasia of the fibrous sheath).
 Human sperm mitochondrial DNA (mtDNA) is a circular, histone- and intron-free DNA ring that
encodes for respiratory-chain-complex proteins and is very susceptible to mutations.

SUMMARY
 Spermatogenesis is a remarkably intricate and complex process that is driven by precisely regulated
and pulsatile secretions of GnRH, LH, and FSH from the hypothalamic-pituitary-gonadal axis.
Pertubations in this hormonal milieu are common causes of male infertility. Sperm production
occurs in the testis, a specialized structure that functions optimally at 2° C to 4° C below body
temperature and generates a mature human sperm in 64 days.
 Well-integrated cycles and waves of spermatogenesis ensure that human sperm production is
constant at about 1200 sperm per second. Spermatogenesis is an androgen-dependent process that
occurs with very high intratesticular levels of testosterone. The product of spermatogenesis, the
spermatozoa, leave the testis as immotile cells with limited capacity to fertilize an oocyte. After
epididymal transit, sperm are typically motile and capable of fertilization.
 During ejaculation, sperm are rapidly transported through the ejaculatory ducts into the urethra
from the distal epididymis. The ejaculate itself supports sperm metabolism, motility, serves as an
antioxidant, and serves as a barrier to exclude subsequent gamete deposits from gaining access to
the egg.

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