Dr.

Niranjan Murthy HL Asst Prof of Physiology ESIC-MC & PGIMSR Bangalore

 Preservation and perpetuation of species

 Asexual reproduction- genes from a single parent Sexual reproduction- fusion of male and female gametes

 Fission- invertebrates

 Budding Fragmentation

 Parthenogenesis fishes, amphibians and lizards Hermaphroditism Sequential hermaphroditism

(a) Both lizards in this photograph are C. uniparens females. The one on top is playing the role of a male. Every two or three weeks during the breeding season, individuals switch sex roles.

Ovary size

Ovulation Estrogen Hormones

Ovulation Progesterone

Time

Figure 46.3a, b

Femalelike

Malelike

Femalelike

Malelike

(b) The sexual behavior of C. uniparens is correlated with the cycle of ovulation mediated by sex hormones. As blood levels of estrogen rise, the ovaries grow, and the lizard behaves like a female. After ovulation, the estrogen level drops abruptly, and the progesterone level rises; these hormone levels correlate with male behavior.

Behavior

 External fertilization

Ovaries

Oviduct

Internal fertilization
Uterus Follicles Uterine wall Endometrium Corpus luteum

Cervix Vagina Seminal vesicle (behind bladder) (Urinary bladder)

Prostate gland Urethra Scrotum Figur Glans penis e 46.1 0 Bulbourethral gland Erectile tissue of penis Vas deferens Epididymis Testis

Physiology of Human Reproduction

Fertilization Parturition Lactation Sexual dimorphism completes at puberty

Sex determination
Gender differences are mainly by Y chromosome and GONADS Gonads have dual function- gametogenic and endocrine Gonadal function depends on gonadotropins Cyclic secretion in females and noncyclic in males

 Chromosomal sex:

 Sex determination is by 1. Karyotyping 2. Sex chromatin

Barr bodies

Development of reproductive system

Development of gonads: Primitive gonad arises from genital ridge Identical in both genders till 6th week Cortex regresses in genetic males in 7-8th week Medulla regresses in genetic females No hormonal effects on gonadal development

Development of genitalia: In 7th week of gestation Mullerian duct in females- develops into uterus and fallopian tubes Wolffian duct in males- develops into epididymis and vas deferens

 External genitalia bipotential till 8th week Urogenital slit closes in males and remain open in females

 Testis secrete MIS and testosterone MIS causes regression of mullerian duct on same side Testosterone induces development of vas deferens and epididymis on same side DHT induces male external genitalia and secondary sexual characters

Brain development: Male behavior and male pattern of gonadotropin secretion if exposed to androgens early

Sex differentiation
1. Genetic sex: Male genetic sex is determined by Y chromosome SRY gene encodes TDF SRY is essential for maleness but not adequate Absence of SRY causes development of ovaries

2. Gonadal sex: Differentiation occurs around 7-8 weeks

3. Genital sex: Testosterone and MIS in males Absence of these factors leads to development of female tract

Abnormalities of sex differentiation

I Chromosomal abnormalities: 1. Turners syndrome: ovarian agenesis 45 XO pattern Dwarfism, webbing of neck, diminished sexual development

2. Klinefelter syndrome: Seminiferous tubule dysgenesis 47 XXY Female genetic sex with male gonadal and genital sex Tall stature Mental retardation

3. Superfemale: 47 XXX No abnormalities 4. True hermaphroditism: XX/XY mosaic Both gonads present

II Hormonal abnormalities: Pseudohermaphroditism- Genetic sex of one type with external genitalia of opposite sex 1. Female Pseudohermaphroditism- Genetic females exposed to androgens by 8th to 13th week 2. Male Pseudohermaphroditism- Genetic males with female genitalia due to defective testis or androgen resistance

Puberty
Transition from the non-reproductive state to the state of reproductive functions that allows procreation Activation of gonads and development of reproductive tract Period of maturation is called ADOLESCENCE

Events of puberty: Thelarche Pubarche Menarche Adrenarche

 Control of onset of puberty Pulsatile secretion of gonadotropins Role of leptin

Sexual precocity

Male Reproductive System

Gonads- Testes Reproductive tract- internal and external genitalia Accessory sex glands- prostate, seminal vesicle, bulbourethral glands Lifelong spermatogenesis

 Testes formed in abdomen Descend into scrotum in 7th month of intrauterine life Requires MIS and other factors Cryptorchidism Slightly lower body temperature [2 C] Gametogenesis and hormonal secretion

Scrotum
Sac of skin and superficial fascia that hangs outside the abdomino-pelvic cavity at the root of the penis Role or cremaster and dartos?? Contains paired testicles separated by a midline septum Its external positioning keeps the testes 2C lower than core body temperature

Testis
Covered by tunica vaginalis and albuginia 300 lobules with 2-4 Seminiferous tubules Each lobule has intratubular compartment (Seminiferous epithelium) and peritubular compartment (neurovascular elements) Seminiferous epithelium- sperm cells and sertoli cells

MALE REPRODUCTIVE SYSTEM

TESTIS

TUNICA ALBUGINEA

EPIDIDYMIS

LOBULES

TESTIS H&E SEMINIFEROUS TUBULES

TESTIS H&E SEMINIFEROUS TUBULES

SEMINIFEROUS TUBULES INTERSTITIAL CONN. TISSUE

 Cells of testis: 1. Germ cells 2. Sertoli cells 3. Leydig cells

Spermatogenesis
Spermatogonia Primary spermatocyte Secondary spermatocyte Spermatid Mature sperm

 Spermatogonia- stem cells- enter meiosis Incomplete cytokinesis Primary spermatocyte- prophase of 1st meiotic division Secondary spermatocyte- completion of 1st meiotic division Spermatids- completion of 2nd meiotic division Spermatozoa- spermiogenesis

 Duration of 72 days Recruitment of adjacent spermatogonia every 16 days Spermiogenesis- maturation of spermatids into spermatozoa Spermiation- release of spermatozoa Further maturation occurs as it passes through the genital tract

Spermiogenesis

Sertoli cells
True epithelium Extend from basal lamina to the lumen From gap junction with each sperm cell stage Tight junction between adjacent sertoli cells forms blood-testis barrier

Functions of sertoli cells

1. 2. 3. 4. 5. 6. 7. 8. 9. Forms blood-testis barrier Provide nutrition to sperm cells Expresses FSH and androgen receptor Express CYP19/aromatase, which converts androgens to estradiol Express ABP Phagocytose residual body Secretes large amount of fluid as a bathing medium for sperms Production of AMH Production of Inhibin

Peritubular compartment
Loose connective tissue Peritubular capillary net work Leydig cells

Leydig cells
Steroidogenic Cholesterol is synthesized de novo, or acquired through LDL and HDL receptors Synthesizes testosterone from androstenedione

Androgens- Fate and Actions

Significant amounts diffuse into seminiferous tubules and get concentrated by ABP 100 times more concentrated than blood Converted to 17 estradiol in sertoli cells Sperm cells have estrogen receptors Aromatase deficiency lead to tall stature Conversion to 5-dihydrotestosterone by 5-reductase

Functions of DHT: Masculinization of external genitalia in utero Growth and activity of prostate Growth of penis Darkening and folding of scrotum Growth of pubic and axillary hair Growth of facial and body hair Increased muscle mass 5-reductase inhibitors- prostatic hypertrophy

Peripheral actions of testosterone: Regulation of sertoli cell function Induces development of male tract Increase LDL and VLDL levels Increased RBC production Promote bone growth Anabolic effect Maintenance of erectile function and libido

Mechanism of action
Cytoplasmic androgen receptors (AR) bound to chaperone proteins ARE- Androgen Response Element Both testosterone and DHT act through AR

Regulation of testicular functions

 Male contraceptive Steroid abuse

Male Reproductive Tract

Epididymis Vas deferens Ejaculatory duct

Accessory structures: Seminal vesicles Prostate Bulbourethral glands Scrotum Penis

Epididymis

Epididymis
Posterior border of testis Coiled and 7mtr long Head, body and tail Sperms spend a month in epididymis and acquire motility Sperm can be stored for several months in tail of the epididymis Decapacitation

Vas deferens

Vas deferens
40-45cm long Ascend as part of spermatic cord Stores sperms Vasectomy

Ejaculatory duct
2cms in length Empties into prostatic urethra

Seminal fluid
Milky white sticky mixture of sperms and glandular secretions Mixture of secretions from glands like prostate, seminal vesicle and bulbourethral glands Provide nutrition to sperms Propulsion of sperm Buffering

Seminal vesicle secretion: Primary source of fructose 60% of seminal fluid Seminogelins- coagulation of semen after ejaculation Prostatic fluid: 30% of volume High in citrate, zinc, spermine and acid phosphatase PSA- liquefies coagulated semen after a few minutes Bulbourethral glands: Rich in mucus Lubrication , cleansing and buffering of urethra

Spermiogenesis
Spermatids lose excess cytoplasm, acquire tail Parts of sperm; Head- contains DNA; covered by acrosomal cap containing enzymes required for penetration of ovum Body- plenty of mitochondriae Tail- flagellum

Journey of spermatozoa
Stays for one month in epididymis and undergoes maturation Decapacitation Stored for several months in tail of epididymis and vas deferens Acquire motility in epididymis by activation of CatSper protein Undergoes capacitation when exposed to female reproductive tract environment

The penis

 Copulatory organ designed to deliver sperms deep into vagina Corpora cavernosa and corpus spongiosum Vascular spaces Circumcision

Erection
Psychic stimuli and impulses from genitalia Efferent parasympathetic nerves: Nervi erigentis Nonadrenergic noncholinergic fibers

Nitric oxide

Guanylyl cyclase

cGMP

Dilation of arterioles

Erectile tissue gets engorged with blood

Veins get compressed

Sympathetic vasoconstrictors to renal arteries

Erectile dysfunction Sildenafil citrate Phosphodiesterase inhibitor Achromatopsia

Emission: propulsion of semen into urethra Ejaculation: ejection of semen out of urethra Afferents- Internal pudendal nerves Emission- Sympathetic response- upper lumbar segments- contraction of smooth muscles of vas deferens and seminal vesicles Ejaculation- contraction of bulbocavernosusS1,2,3 motor

Female Reproductive System

Ovaries Oviducts, Uterus, Cervix, Vagina External genitalia

The Ovary
Bilateral Located in broad ligament Cortex- cellular Medulla- neurovascular elements Ovarian follicles- oocyte surrounded by follicular cells Cortex covered by tunica albuginea and surface epithelial cells

 Functions: 1. Gametogenesis 2. Secretion of female sex hormones 3. Production of Inhibin

Uterus
Provide mechanical support, protection and nutrition to developing conceptus Parts- Fundus, Body and Isthmus

Uterine wall
Endometrium Myometrium Perimetrium

Endometrium 10% of wall thickness Glands and blood vessels Functional and Basilar zone Myometrium 90% Longitudinal, circular and oblique muscles

Cervix
Considered separately from uterus Internal os, cervical canal, external os Extends from isthmus to vagina and projects into it

Fallopian tube
13 cm long Transport gametes Site of fertilization Infundibulum, Ampulla, Isthmus

Vagina
7.5 to 9cm Highly distensible Fornix is a shallow recess surrounding cervical protrusion Hymen- elastic epithelial fold

External genitalia
Vulva or Pudendum Vestibule- area surrounded by labia minora; has urethral and vaginal openings Clitoris- equivalent of penis Labia majora

Menstrual cycle
Ovarian cycle Endometrial cycle Cervical changes Vaginal changes Hormonal changes Usually around 28 days Start from the first day of menstruation

Oogenesis
1. 2. 3. 4. 5. 6. 7. Resting primordial follicle Growing preantral follicle Growing antral follicle Dominant follicle Dominant follicle within periovulatory period Corpus luteum Atretic follicle

Resting primordial follicle

Appear during mid-gestation Made of oogonia and somatocytes Ovarian reserve 7 million ooogonia enter process of 1st meiotic division at 5th month- primary oocytes Primary oocytes get surrounded by follicle cells Follicular reserve: 3lakh at maturity <1000 at menopause

 Small subset of follicles enter growth phase in waves Independent of gonadotropins Gamete- primary oocyte- maturational incompetence- arrest in diplotene stage of 1st meiotic division

Growing preantral follicle

Appearance of cuboidal granulosa cellsprimary follicle Granulosa cells become stratified- secondary follicle Formation of thecal cells- mature preantral follicle Move inwards and release angiogenic factors

Gametes secrete ZP1, ZP2 & ZP3. Form zona pellucida- species specific Granulosa cells express FSH receptor Thecal cells secrete small amounts of androstenidione and express LH receptor

Growing antral follicles

25 days to grow into early antral follicle Granulosa cell become 6-7 layered Formation of antrum Reach 2-5mm in 45days Recruitable follicle Stratum granulosa- steroidogenic Cumulus oophorus Highly dependent on FSH

 Oocyte acquires maturation competence but still maintains meiotic arrest Maintenance of high levels of cAMP Thecal cells produce increased amounts of androgens FSH stimulates expression of aromatase in granulosa cells Inhibin B formation by granulosa cells

Dominant follicle
About 20 follicles are recruited Prolifera quota Dominant follicle 20mm in preovulatory phase Oocyte becomes 140 in diameter Dominant follicle- significantly steroidogenic FSH induces LH receptors In mural granulosa cells

Dominant follicle during periovulatory period

From onset LH surge to ovulation 32-36hrs Luteinization process

Ovulation
LH surge Oocyte progresses to metaphase of meiosis II Formation of stigma Release of cytokines and hydrolytic enzymes Breakdown at stigma Cumulus-oocyte complex released into peritoneum

Ovulation
Cumulus expansion- hyaluronic acid and matrix components Granulosa cells secrete angiogenic factors

Corpus luteum
Corpus hemorhagicum Enlargement of granulosa lutein cells 14 days life Needs hCG PGF2 cause regression of lutein cells Corpus albicans Peaking of progesterone production Inhibin A repress FSH

 Fall in hormones during 2nd half of luteal phase Luteal phase deficiency

Atretic follicles
Apoptosis Thecal cells persist

Ovarian cycle
Follicular phase: preovulatory phase Luteal phase: postovulatory phase Luteal phase is fixed- 14 days

Uterine cycle
Proliferatory phase: follicular phase Secretory phase: luteal phase Proliferatory phase begins at day 1 of menstruation Menstruation: process of endometrial (stratum functionale) sloughing; lasts 1-5 days; loss of upto 80ml (average 35ml) of blood; withdrawal of progesterone

Proliferative phase: begins on 1st day of menses Till ovulation Increase in endometrial thickness Endometrial glands are drawn out Influence of estrogen Estrogen induces progesterone receptor expression

Secretory phase: Glands get coiled & tortuous and secrete clear fluid Under the influence of progesterone Change in adhesivity of surface epithelium Differentiation of stromal cells into predecidual cells

Menopause
45-55yrs Decline in number of follicles Reduced estrogen and progesterone High gonadotropin levels Climacteric Hot flushes coincide with LH surge

Disorders of menstruation
Menorrhagia: excessive bleeding Metrorrhagia: irregular and intermenstrual bleeding Dysmenorrhoea: painful bleeding Oligomenorrhoea: Amenorrhoea: Anovulatory cycle: Endometriosis: cause of infertility

Cyclic changes in uterine cervix

Estrogen makes cervical mucus thinner and alkaline Progesterone makes it thick, tenaceous, acidic and cellular Spinnbarkeit and fern pattern in ovulatory phase

Vaginal cycle
Estrogens cornify vaginal epithelium Progesterone causes proliferation of vaginal epithelium and infilteration with leucocytes Thick mucus secretion due to progesterone Cornification- increased glycogen accumulation

Hormonal regulation of menstrual cycle

Ovarian hormones
Estrogens progesterone

Estrogens

 LH acts on theca interna to increase androgen secretion FSH induces aromatase in granulosa cells Androgens are converted to 17 estradiol 2% of circulating estrogen is free; 60% bound to albumin; rest to SHBG Metabolized in liver

Actions of estrogens
1. 2. 3. 4. 5. 6. 7. 8. Facilitate growth of follicles Increase motility of fallopian tubes Support endometrial growth Increase myometrial excitability Feedback inhibition of FSH Dual effect on LH regulation Increase libido Increased dendrites and synaptic knobs in rat neurons

8. Reported to slow progression of alzheimers 9. Growth hormone of breast 10. Secondary sexual character development 11. Salt and water retention 12. Lowers plasma cholesterol 13. Anti-atherogenic 14. Avoids osteoporosis

Mechanism of action: Nuclear receptors ER and Er Evidence of surface receptors

Progesterone
Produced by granulosa cells Stimulated by LH

 2% is freely circulating; 80% bound to albumin and remaining to CBG Converted to pregnenidiol and conjugated

Actions of progesterone
1. Secretory changes in endometrium. Prepares it for maintenance of conceptus 2. Decreases the excitability of myometrium 3. Decrease estrogen receptors and increase conversion of active estrogens to less active ones 4. Stimulate development of lobules and alveoli of breast

5. Thermogenic 6. Stimulate respiration 7. Natriuresis in large doses

Mechanism of action: Nuclear receptor bound to HSP Mifepristone

Control of ovarian function

GnRH secreted in episodic bursts Continuous infusion of GnRH causes downregulation of receptors in pituitary GnRH frequency increases by estrogen and decreases by progesterone Self priming of GnRH in mid-cycle Constantly elevated levels of GnRH will downregulate its receptors in pituitary: treatment of precocious puberty and prostatic cancer

 Constant elevation of estrogen to 300% (200pg/mL) for 36hrs induces LH surge PGF2 luteolysin ET-1 and oxytocin

Tests for ovulation

1. 2. 3. 4. Increase in BBT Ultrasonography Cervical smear Spinnbarkeit

Changes during intercourse

VIP- vasodilation- increased blood supply Stimulation of labia, clitoris, nipples Increased vaginal secretion Orgasm

Fertilization
Recombination of genetic material Initiation of embryonic development Ampulla or junction of ampulla and isthmus

 Steps of fertilization: 1. Penetration of cumulus- hyaluronidase 2. Bind to ZP3- Acrosomal reaction- ZP2 binding 3. Fusion of membranes 4. Ca2+ cascade- egg activation 5. Exocytosis of cortical granules- modify ZP 2 & 3 6. Sperm enters ovum- completion of 2nd meiotic division

Implantation
Fertilization on day 16 Implantation after 6 days Morula- 3days Blastocyst 4-5 days Inner cell mass and trophoblast Embryonic pole Digest ZP by proteases 5-6 days: hatched blastocyst

 Cytotrophoblast Syncytiotrophoblast- adhesive; invasive; endocrine; phagocytosis; transport of substances Interstitial implantation hCG and other hormones

Maternal response: Decidualization Tissue inhibitors of metalloproteinases

Failure to reject fetus

Placental trophoblast express HLA-G Fas ligand

Human chorionic gonadotropin

Glycoprotein Trophoblastic cells, blastocyst Peaks in 60-90 days Similar to LH, FSH and TSH Luteotrophic Marker of pregnancy

Human chorionic somatomammotropin

Maternal GH of pregnancy Proportional to the size of placenta Retention of calcium, nitrogen and potassium Lipolysis Reduced glucose utilization

Other hormones
Relaxin CRH, -endorphin, -MSH GnRH, Inhibin subunit of hCG, Prolactin

Fetoplacental unit
Placenta takes over in 6th week after pregnancy Placenta synthesize progesterone and pregnenolone Progesterone is converted to cortisol in fetal adrenal Pregnenolone forms DHEAS & 16OH-DHEAS in fetal liver They form estradiol and estriol respectively in placenta

Functions of estrogen in pregnancy

Growth of uterus Increase blood flow to fetus Growth of breast Increased size of maternal external genitalia

Functions of progesterone in pregnancy

Decidualization Reduces excitability of myometrium Increases secretions of tubes and uterus before implantation Growth of alveoli of breasts

Placenta
Formed by trophoblastic cells Bathed with maternal blood 2 umbilical arteries and one vein

Functions of placenta
Fetal nutrition Supply oxygen and remove CO2 Remove metabolic wastes Hormonal production Maintenance of pregnancy via progesterone

Maternal changes during pregnancy

Weight gain-12-14 kgs; 3.5kg fetus; 2kg amniotic fluid; 2kg uterus; 2kg breast, 3kg ECF; 1.5kg fat BMR increases by 15% Cardiac output increases by 30-40% at 27th week Blood volume increases by 30% Dilutional anemia Neutrophilia, increased procoagulant activity Increased RR and minute ventilation

 GFR increases by 50% Increased renal reabsorption of sodium and water Relaxation of ligaments Melasma gravidarum; striae gravidarum Reduced IOP; subconjunctival hemorrhages; corneal edema Nasal congestion; hoarseness of voice Increased salivation

 Endocrine and metabolic changes: Thyroid enlargement; Increased Iodine uptake; Increased thyroglobulin Reduced serum Ca2+ levels; increased Vit D; intestinal absorption of calcium Increased pituitary size Hyperplasia and hypertrophy of cells of langerhans; increased glycogen storage

Amniotic fluid
0.5 to 1.0 ltr Contributed by fetal urine Replaced every 3hrs Amniocentesis

Parturition
Duration of pregnancy- 284 days from LMP Increased CRH secretion- increased cortisollung maturation Increase in estrogen & fall in progesterone Increased PGF2 Increase in oxytocin receptors in myometrium Parturition reflex Bearing down

Parturition reflex

Tests for Pregnancy

I Biological tests: 1. Ascheim Zondek test- urine of pregnant injected to female mice 2. Friedmans test- pregnant urine injected into female rabbit II Immunologic tests: Antibody prepared from rabbit Urine + anti-HCG + HCG coated sheep RBCs