The Thyroid

Thyroid Gland Located on the front and sides of the trachea just below the larynx, 30 grams, highly vascularized Hormones: o T4 and T3 Target organ is all tissue Stimulates energy production and protein synthesis which contribute to the overall growth of the body o Calcitonin Lowers the blood calcium levels Target organs are bones, kidneys and intestines Helps maintain normal levels of calcium in blood and maintain stable strong bones Functional units = follicles ~3 million follicles/thyroid follicle epithelial cells produce thyroid hormones (T3 and T4) clear cells (C cells) produce calcitonin the endocrine function of the thyroid gland is regulated by sympathetic nervous system

Thyroid Development most of the thyroid arises from the primitive GI tract (endoderm) but thyroid c cells come from the neural crest (ectoderm) thyroid is often asymmetric (right bigger than left) female thyroid is bigger than the male thyroid (increases during puberty, pregnancy lactation and season) o at these times the body requires more thyroid hormone so it increases its size to produce more hormone

Thyroid Hormones thyroid hormones are the only substances in the body that have iodine in their structure the main source of iodine is through dietary iodide including iodated bread, iodized salt and dairy products

Thyroid Hormones in Circulation strongly bound to serum thyroid binding proteins .03% of T4 is free and .3% of T3 is free thyroid binding globulin (TBG) = high affinity transthyretin = moderate affinity serum albumin = low affinity serum half life of T4 is about 7 days serum half life of T3 is about 1 day The Iodide Transporter How does iodide get into the thyroid follicle cells? Sodium/iodide symporter protein (NIS) o Responsible for transport of iodide from the blood into epithelial cells o A transmembrane span protein o Thyroid follicle cells sequester iodide against a 30X gradient mainly by NIS (iodide is 30X higher in the epithelial cells than in the blood) o The thyroid concentrates inorganic iodide from the extracellular fluid by NIS o NIS I stimulated by thyroid releasing hormone TSH o NIS is inhibited by an increasing grandular content of organic iodine o NIS activity is sodium dependent

How does iodide get into the follicular lumen from the thyroid follicle? Pendrin protein o A 11 transmembrane span protein o Transports chloride, iodide and bicarbonate o Its activity is sodium dependent o Pendrin is positively regulated by the autocrine action of thyroglobulin

Thyroid Hormone Synthesis Precursor amino acids: o Diiodotyrosine (DIT) o Monoiodotyrosine (MIT)

o DIT and MIT are formed by the iodination of tyrosine residues within the matrix of thyroglobulin Thyroglobulin o Large glycoprotein o ~10% carbohydrate o ~1% iodide o composed of two weakly associated identical subunits o ~125 tyrosine residues (1/3 are available for reaction) o unique protein to the thyroid o synthesized in the thyroid follicle cells and secreted into the follicle lumen o both T3 and T4 form within thyroglobulin

Synthesis of T4 on Thyroglobulin

MIT + DIT = T3 DIT + DIT = T4

Thyroid Hormone Metabolism T4 is the predominant secretary product of the thyroid, the thyroid normally makes much more T4 than T3 if iodide intake is adequate and the T4 to T3 ratio of normal thyroglobulin is 15:1 T4 is peripherally converted to receptor-actibve T3 by outer ring deiodination (ORD) to inactive reverse T3 (rT3) by inner ring deiodination 5 deiodinase converts T4 to T3 at target tissues hepatic type 1 deiodinase (D1) is responsible for most of the T3 in the circulation (most of the T3 in circulation comes from the liver) reverse T3 is an inactive metabolite thyroid mainly produces T4 not T3 but T3 is the major biologically active, most potent form of thyroid hormone

Actions of Thyroid Hormones in Mammals Hormone Homeostatic Control o Anterior pituitary negative feedback regulation of TSH release o Required for pituitary synthesis of PRL and GH Growth and Development (as tissue growth factors) o Regulates growth (with GH) o Required for early neural development o Required for bone growth and development Increases basal metabolic rate (BMR) or calorigenesis o Brain-stimulates feeding behavior o Increases oxygen consumption o Increases heart rate o Decreases liver glycogen o Decreases plasma cholesterol o Required for liver conversion of carotenes to vitamin A o Thermogenesis and temperature regulation- increases mitochondrial oxygen consumption and ATP synthesis o Induces synthesis of many enzymes and other cellular proteins

Thyroid Stimulating Hormone (TSH) Roles of TSH: Affects thyroid gland activity via stimulation of the TSH receptor cAMP pathway Stimulate thyroid hormone biosynthesis and release o Activates iodide uptake by thyroid o Activates T3. T4 synthesis on thyroglobulin (TG) in the thyroid colloid o Activates TG-T3.T4 uptake into the thyroid epithelial cells o Activates lysosomal release of T3 and T4 from TG o Activates the release of T3 and T4 into the blood Promotes general growth of thyroid gland

Thyroid Hormone Receptor Signaling Thyroid hormone receptors are nuclear receptors Predominantly work as heterodimers with RXR May be mostly nuclear even in the absence of the ligand 3 major isoforms of the thyroid receptor

Thyroid Hormone receptor Signaling

Target Gene Example for Thyroid Hormone

Thyroid Diseases Hypothyroidism Deficiency of thyroid hormones (hypothyroxinemia) common disease (1 in 4000) Primary Hypothyroidism o Thyroid gland resistance o Cretinism = childhood hypothyroidism Goitrous cretinism (lack of iodine) Athyreotic cretinism (congenital absence if thyroid) o Myxedema = adult hypothyroidism Simple goiter (lack of iodide) Idiopathic (thyroiditis of unknown cause) Iatrogenic (surgical removal or chemical inactivation) Spontaneous (autoimmune disorder; Hashimotos thyroiditis) Secondary Hypothyroidism o Pituitary defect o Hypothalamic hypothyroidism Familial end organ resistance to thyroid hormones Possible clinical symptoms of hypothyroidism o Appearance: weight gain, possible goiter, puffy appearance, loss of hair o Low BMR: low body temperature, decreased perspiration o Disposition: lethargic, depressed, intolerant to cold

Examples of Molecular Defects Underlying Hypothyroidism Mutations in the NIS gene o Cause hypothyroidism Mutations in the Thyroid Peroxidase (TPO) gene o Cause hypothyroidism o No organification of iodide Mutations in the Thyroglobulin (TG) gene o Cause hypothyroidism Mutations in the TR beta gene o Cause thyroid hormone resistance

Hyperthyroidism Excess of thyroid hormones (thyrotoxicosis) Common disease (2% adult females) Primary Hyperthyroidism o Adenoma a noncancerous tumor of the glands o Carcinoma a tumor of the epithelial cells Secondary Hyperthyroidism o Pituitary defect, e.g. excess TSH o Hypothalamic hyperthyroidism, e.g. excess TRH o Pituitary thryotrope unresponsiveness to T3 feedback inhibition o Ectopic TSH secretion o Long acting thyroid stimulator (LATS ) (Graves Disease) Autoimmune Disease Antibodies to the TSH-R are produced Thyroid is always on Feedback inhibition loop lowers TSH , but thyroid still active

Possible clinical symptoms of hyperthyroidism o Appearance: weight loss and possible goiter o High BMR, high body temperature o Disposition: agitated, nervous, easy to fatigue, intolerant to heat, sweating o Cardiac function: tachycardia, atrial fibrillation o Loss of muscle mass

Thyroid Cancer An easily cured form of cancer with surgery and radioactive iodine Various forms of thyroid cancer affect an estimated 14,900 women and 4600 men every year Thyroid Goiter Can find with hypo/hyperthyroidism Graves Disease Clinical presentation of Graves ophthalmopathy o Retraction of upper eyelids o Severe periorbital edema o Predominanty unilateral movements o Spontaneous subluxaton of a severely proptotic left eye

1. Normal 2. Thyroid gland cant produce enough T3, T4 so no feedback TSH increases 3. Hypothalamic pituitary problem so impair TSH production so decrease T3, T4 4. Problem id hypothalamus or pituitary response to T3, T4 often T3 receptor mutation, target tissue dont respond to T3, T4 5. Thyroid produces too much T3, T4 so it functions to decrease TSH and T3, T4 higher 6. To much TSH so thyroid is stimulated to over produce T3 and T4 so both higher 7. Autoimmune problem, autoimmunity produced by different tissue too much T3, T4 in blood, TSH production decreased Parathyroid Gland There are four parathyroid glands: two on the back of each lobe of the thyroid gland Hormone o Parathyroid hormone Raises blood calcium levels An antagonist to calcitonin

Reproductive Endocrinology I Sexual Differentiation The Paradigm for Sexual Differentiation

Sex diffferentiation is a sequential process that begins at fertilization with the establishment of chromosomal sex, continues with the determination of gonadal sex and culminates in the development of secondary sexual characteristics that comprise the male and female phenotypes (phenotypic sex)

Development of Chromosomal Sex Normal genotypes of human germ cells in the haploid state: Oocytes: 23, X (22 autosomes, 1 X chromosome) Spermatocytes: 23, X or 23, Y (22 autosomes, 1 X or 1 Y chromosome) Normal genotypes of human somatic cells in the diploid state: Females: 46, XX Males: $6, XY (22 pairs of autosomes, pair of sex chromosomes) Chromosomal sex depends on the presence or absence of the Y chromosome The Y Chromosome One of the smallest human chromosomes Contains more than 30 genes Contributes to functions including spermatogenesis, skeletal growth and tooth development Contains psuedoautosomal regions as its ends that permit the Y chromosome to pair with the X chromosome during meiosis The most striking function of the Y chromosome is in sex determination

Genetic Regulation of Sexual Development The SRY gene SRY = Sex-determining Region, Y chromosome The SRY gene is inside the pseudoautosomal region of the Y chromosome The SRY gene encodes a transcription factor that triggers a cascade of events that result in the development of testes. Testis development depends on inheritance of this Y chromosome encoded TDF, testes determining factor
SRY product 22 Y 22 X 44 XY testis

testosterone male phenotype

22 X mature sperm

22 X mature ova

44 XX zygotes

ovary

female phenotype

The SRY gene is both necessary and sufficient for male sex determination, 44 44 44 44 functioning as a XO switch. In presence of Y, gonad follows the testis sex XXX YO XXY pathway. In absence of Y chromosome, gonads follow the ovarian pathway gonadal super Some sex reversal patients have SRY gene lethal mutations seminiferous
dys genesis female in utero tubule dys genesis

Development of Gonadal Sex In the first 7-8 weeks of gestation, the sex of the embryo cannot be recognized indifferent phase During the indifferent phase, the gonads are identical in both sexes indifferent gonads Differentiation of Internal Genitalia of the Human Fetus o During the indifferent phase, the embryo acquires dual ductal systems that are precurs to male and female internal genitalia o The Wolffian (or mesonephric) duct: give ris eti the internal male accessory organs of reproduction the seminal vesicles, vas deferens and epididymis o The Mullerian (or paramesonephric) duct: gives rise ti the internal female reproductive tract (fallopian tubes, uterus and upper vagina) Differentiation of External Genitalia of the Human Fetus o During the indifferent phase the embryo acquires genital tubercle, genital fold and genital swelling that are precurs to the male and female external genitalia

Development of Phenotypic Sex Endocrine Control The fetal testes are necessary for male phenotypic development. Two hormones of the fetal testis are essential. o AMH: anti-Mullerian hormone (product of Sertoli cells) o Testosterone (product of Leydig cells) The fetal ovaries are not necessary for female phenotypic development.

o If remove ovaries and testis from the developing fetus, the Mllerian ducts develop and the Wolffian ducts atrophy, suggesting that the female is the default pathway and that the gonads are not required for development of a female ductal system. Female phenotypic development occurs in the absence of gonads. AMH o ~70 kDa glycoprotein o Found as a dimer (a member of the TGFb superfamily) o Produced by the Sertoli cells of embryonic testes o Causes atrophy (regression) of the Mllerian ducts o In tandem, the Leydig cells of embryonic testes produce testosterone which promotes development of the Wolffian Ducts o In adults made by testes (and ovaries - why?) o Mllerian duct is only sensitive to AMH during an early developmental period (over by 8 wks in humans) - therefore production of AMH in the adult has no deleterious effects o Mutations in AMH associated with PMDS (Persistent Mllerian Duct Syndrome)

Androgens and Androgen Receptor Androgens: Testosterone Dihydrotestosterone Testosterone Dihydrotestosterone

Androgen Receptors A member of the nuclear receptor Bind testosterone and dihydrotestosterone A large group of hydrophobic ligands: steroid hormones, thyroid hormones, retinoids, vitamin D, bile acids, and oxysterols Regulation of Male Sexual Development

Phenotypic Sex: Brain Sex Brain is inherently female, unless androgens direct it to a male fate. Sex hormones may differentiate the human brain from about 6 weeks of age. Androgens alter development of brain especially hypothalamus - these regions mediate sexual behavior & gonadotropin secretion. Male brain: mounting behavior, no cyclic release of FSH and LH. Androgens act through local conversion to estrogens (aromatase). Social effects also clearly important in perception of gender in humans.

Secondary Sex Characteristics Testosterone or DHT actions at puberty in vertebrates Sexual maturation - courtship, aggression, display Sex organs Penis and scrotum enlarge and change character Skin Acne is related to increased sebaceous gland activity Pubic and facial hair Recession of male hairline Skin, hair and feather colorization: androgen dependent pigmentation.

Kidney Gender-dependent difference in mitochondria and lysosomes structures in kidney. Mitochondrial enzymes have higher activities in males. Submaxillary glands in mice Produce NGF and EGF under androgen control. Genetic Regulation of Sexual Development SRY: Sex-determining Region, Y chromosome AMH: Anti-Mllerian Hormone SF-1: Steroidogenic Factor 1 o an orphan nuclear receptor (transcription factor). o Regulates steroidogenesis. Activates the expression of genes involved in steroidogenesis (e.g. P450 steroid hydroxylases, 3-hydroxysteroid dehydrogenase, ACTH receptor, StAR, and high-density lipoprotein receptor) o Stimulate testosterone production. o Stimulates AMH production by activating AMH gene promoter. o Activate the Dax-1 gene promoter. o Mice with SF-1 k/o die at ~p8 with absence of adrenals and gonads SOX9 o Member of a family of SRY-related proteins (Sox = SRY type HMG box proteins). o A transcription factor. o Stimulates AMH production by activating AMH transcription. o important at early stages of sex differentiation. o Mutations in this gene cause broad developmental defects including skeletal problems e.g. campomelic dysplasia (CD). Some CD patients are 46XY females with genital defects. DAX-1 o A nuclear receptor-related protein o A transcription factor o Inhibits the expression of SF-1 target genes WT-1 o A transcription factor associated with Wilms tumor (an embryonic kidney tumor) . o mice with Wt-1 k/o die at midgestation with absence of kidneys and gonads. o Mutations in this gene cause broad developmental diseases (including Wilm's tumor of the kidney, genito-urinary problems, mental retardation, aniridia).

Disorders of Chromosomal Sex Turner Syndrome 45, X genotype, female. Gonads do not develop (gonadal dysgenesis). accounts for up to 10% of spontaneous abortions. ~ 2% of all human conceptuses are 45, X, less than 1% of which survive to term. External genitalia are female. deficiency of ovarian steroid hormones. short stature, webbed neck. the most common cause of primary amenorrhea.

Klinefelter Syndrome Typical genotype : 47, XXY o Atypical genotypes (Klinefelter's variant) : more than 2 X , 1 Y (e.g. 48, XXXY ) predominant phenotype: male ~ 0.2% male births

testes develop, but after birth germ cells degenerate small testes, low testosterone levels failure of puberty, no sperm Impaired virilization and enhanced feminization (breast enlargement) due to an imbalance in the estrogen/androgen ratio

Disorders of Gonadal Sex Chromosomal sex is normal. Differentiation of the gonads is abnormal. Phenotypic sex varies, depending on when the gonadal defect becomes manifest during embryogenesis. 46, XY Complete Gonadal Dysgenesis Mutations of genes implicated in complete gonadal dysgenesis o SRY mutation: 10-20% of the patients o SF-1 mutation o SOX9 mutation o DAX-1 mutation o WT1 mutation The sex phenotype is female because gonadal development is arrested before AMH and androgens are produced.

Disorders of Phenotypic Sex The phenotypic sex is ambiguous or is completely in disagreement with the chromosomal and gonadal sex. Results from a failure of synthesis or action of hormones that mediate male sexual differentiation---Male Pseudohermaphroditism. Results from the inappropriate synthesis of androgensFemale Pseudohermaphroditism. Pseudohermaphroditism - (congenital) condition in which a person has external genitalia of one sex and internal sex organs of the other sex. Male Pseudohermaphroditism 46, XY genotype have testes, but external genitalia and phenotype are female. defects in androgen synthesis in the testis insufficient androgen production in the testes and adrenals. o e.g. StAR mutation, 17-hydroxylase deficiency defects in androgen action in target cells -- androgen resistance due to mutation of AR. defects in Mullerian duct regression - Persistent Mllerian Duct Syndrome (PMDS) o e.g. MIS mutations/deficiency or MIS receptor defects

Androgen Receptor Defects Complete androgen insensitivity syndrome (CAIS) Partial androgen insensitivity syndrome (PAIS) AR on the X chromosome Male pseudohermaphroditism Female external genitalia (phenotype) Normal testes produce MIS - therefore no Mllerian-derived structures vagina is a blind pouch ~1:40,000 frequency Female Psuedohermaphroditism 46, XX genotype have ovaries, but external genitalia are male. can be caused by enzyme deficiencies-- The most common cause of virilization in newborn females is Congenital Adrenal Hyperplasia (CAH) o e.g. 21-hydroxylase (CYP21)deficiency o e.g. 3-b-hydroxystroid dehydrogenase deficiency

Propecia (finasteride) Finasteride is a specific inhibitor of steroid Type II 5a-reductase, an intracellular enzyme that converts the androgen testosterone into 5dihydrotestosterone (DHT). Inhibition of Type II 5a-reductase blocks the peripheral conversion of testosterone to DHT, resulting in significant decreases in serum and tissue DHT concentrations. Contraindications: o Pregnancy. Finasteride use is contraindicated in women when they are or may potentially be pregnant. Because of the ability of 5a-reductase inhibitors to inhibit the conversion of testosterone to DHT, finasteride may cause abnormalities of the external genitalia of a male fetus of a pregnant woman who receives finasteride.

Reproductive Endocrinology II Female Reproductive Endocrinology

Structural Organization of the Ovary

Outer cortex: contains follicles, most of the mass of the ovary Inner medulla: contains stromal cells, and steroid producing cells Hilum: the point of entry of the nerves and blood vessels; the attachment region to the mesovarium Ovary follicles o Functional units of the ovary o Present in different states of developmental or degeneration o Each encloses an oocyte surrounded by granulosa cells and thecal cells

Ovary Hormones o Steroid Hormones Progesterone Estrogens estradiol estrone Androgens androstenedione testosterone o Peptide hormones Relaxin Inhibins Activins Proopiomelanocorticotropin (POMC) family Adrenocorticotropin (ACTH) Alpha-melanocyte-stimulating hormone (alpha-MSH) Beta-endophin Vasopressin Oxytocin

Biosynthesis of Steroids in the Ovary

Pituitary Control of Ovarian Hormone Formation ---- Follicular Two-cell, two gonadotropin pathways

Steroid Hormone Transports and Metabolism Most circulating estrogens (~70%) bind to blood transport proteins, preferentially albumin with a low affinity, or sex hormone-binding globulin (SHBG) with a higher affinity. Estrogens are metabolized predominantly in the liver via oxidation, glucuronidation, and sulfation, and are excreted in the urine. Progesterone circulates bound to corticosteroid-binding protein (transcortin). Progesterone is metabolized in the liver via glucuronidation, and is excreted in the urine.

Actions of Ovarian Steroids Intraovarian Actionsfollicular development Estradiol (E2) o induces proliferation of the granulosa cells; o increases E2 receptors in the granulosa cells; o facilitate the action of FSH in the granulosa cells. Progesterone o inhibits the action of FSH in the granulosa cells; o stimulates ovulation by inducing proteolytic enzymes.

Androgens o enhance the action of FSH in the granulosa cells.

Extraovarian Actions ---- hypothalamic-pituitary axis, reproductive tissues, nonreproductive tissues. Estradiol (E2) o induces proliferation of the endometrium of the uterus; o stimulates the activity of uterine endocervical glands (mucus production); o induces both proliferation and keratinization of the epithelium of the vagina; o stimulates breast development and growth; o increases the amount of plasma proteins produced by the liver; o increases the circulating levels of high-density lipoproteins (HDL), very-lowo density lipoproteins (VLDL), and triglycerides; o lowers the circulating levels of cholesterol and low-density lipoprotein (LDL); o stimulates bone growth and prevents bone resorption (osteoporosis); o enhances the reabsorption of sodium in the kidney. Progesterone o inhibits the proliferation of the endometrium; o suppresses the activity of uterine endocervical glands (mucus production) o inhibits both proliferation and keratinization of the epithelium of the vagina; o increases the circulating levels of cholesterol and LDL and decreases HDL;

Estrogen Receptors Estrogen signals through estrogen receptors (ERs). ERs act in a similar fashion to GR (heat shock protein complexes, etc.). Two isoforms: ER alpha and ER beta. ER alpha and beta are widely co-expressed in different tissues. ER alpha and beta can form homodimer or heterodimer. o Therefore alpha, beta, and alpha/beta pathways The expression levels of ER alpha and beta are different in some tissues. Pituitary > Prostate > CNS > Lung > Bone >

Breast Cancer > cells E2 binds equally well to both alpha and beta receptors. Estrone binds preferentially to alpha receptor. Some environmental estrogens bind preferentially to beta receptor. Estrogen Receptor alpha knockout mouse o Appear normal o Normal sex ratio o But infertile - both sexes o No functional corpora lutea in females o Sperm count low in males o Breast development is poor o Bone density is low (osteoporosis) ERbeta Knockout Mice o Exhibited phenotypes distinct from ER-alpha knockout mice. o ER -/- mice developed normally. o ER -/- females are fertile and exhibit normal sexual behavior, but have fewer and smaller litters than wild type mice. o Reduced ovarian efficiency. o Mutant females had normal breast development and lactated normally. o Young mutant male mice showed no overt abnormalities and reproduced normally. o Older mutant males displayed signs of prostate and bladder hyperplasia. o These results indicate that ER is essential for normal ovulation efficiency but is not essential for female or male sexual differentiation, fertility, or lactation. ERalpha and beta Double Knockout Mice o Both sexes exhibit normal reproductive tract development but are infertile. o Ovaries of adult females exhibit follicle transdifferentiation to structures resembling seminiferous tubules of the testis, including Sertoli-like cells and expression of Mullerian inhibiting substance, sulfated glycoprotein-2, and Sox9. o Therefore, loss of both receptors leads to an ovarian phenotype that is distinct from that of the individual ERKO mutants, which indicates that both receptors are required for the maintenance of germ and somatic cells in the postnatal ovary.

Progesterone Receptor Corpus luteum is major circulating source of progesterone. Progesterone signals through the progesterone receptor (PR). PR acts in similar fashion to GR (heat shock protein complexes, etc.) Progesterone Receptor KO Mice o Development of embryos to adulthood at normal Mendelian frequency with no deviation in the sex ratio. o Impairment in the induction of a sexual behavioral response. o Males are fertile, females infertile. o Extensive reproductive abnormalities in female: Lack of ovulation Uterine hyperplasia and inflammation Severely limited mammary gland development LH and FSH The HPG axis: hypothalamus - pituitary - gonads FSH and LH are the gonadotropins Made in gonadotrope cells of the AP (~10-15% of AP cells) Glycoprotein dimer hormones (like TSH) Luteinizing hormone = LH o In female induces ovulation o In female initiates steroidogenesis in ovarian follicle o In female maintains secretory functions of corpus luteum o In male stimulates Leydig cells to produce testosterone o Half-life = 60 mins Follicle-stimulating hormone = FSH o In female stimulates development of ovarian follicles o In female stimulates secretion of estrogen o In male stimulates spermatogenesis o In male stimulates sex hormone-binding globulin o Half-life = 170 mins

Activins and Inhibins Activin activates and inhibin inhibits pituitary follicle-stimulating hormone (FSH) production. Activin is produced by the testes, ovaries, brain, pituitary gland and erythropoietic tissues. Inhibin is produced by the testes and the ovaries, also in brain, pituitary, adrenal, spleen, placenta, bone marrow, and kidney. They are dimers formed by different combinations of one alpha subunit and two beta subunits. The subunits are joined by disulfide bonds.

Activin and Inhibin Proteins Members of the TGFbeta (transforming growth factor beta) superfamily of growth and differentiation factors (>30 members). At least three subgroups: TGF family: TGFbeta1, TGFbeta2, TGFbeta3 Activin family: Activin A, Activin B, Activin AB, Inhibin A, Inhibin B DVR family: DPP, Vg-1, BMP-4, GDNF, GDFs, Nodal, MIS Activin in Mammals activates FSH production regulates hormone production in the placenta induces differentiation in erythroblasts induces differentiation in osteoblasts promotes survival of neural cells - neurotrophic

inhibits proliferation of gonadal cells inhibits proliferation of endothelial cells inhibits proliferation of lung epithelial cells inhibits proliferation of hepatocytes more roles uncovered frequently

Activin and Inhibin Binding Proteins Follistatin o glycosylated monomeric protein o suppresses pituitary FSH release FSH antagonist o higher affinity for activin than inhibin o regulator of activin bioavailability Alpha 2-macroglobulin o high mol wt tetrameric protein o binds many cytokines and growth factors o broad spectrum protease inhibitor o role unclear Both are produced in hormonally-regulated fashion by gonads

Activin Receptors Transmembrane serine/threonine kinases Type I and Type II subfamilies o Type I receptors: ActR-I, ActR-IB, TSR-I o Type II receptors: ActR-II, ActR-IIB

o Type II receptors bind ligand on their own o Type I receptors need Type II receptors to bind ligand o Both receptor types needed for activin signalling may be shared with other TGFbeta family members e.g. BMP-7 can interact with ActR-I and ActR-II as well as BMPR-I and BMPRII.

Activin Signaling The Smads Pathway

Inhibitory Smads (Smad 6,7) can interfere by binding to the receptors and preventing the phosphorylation and signalling of the pathwayspecific Smads.

Mouse Models to Assess Functions of inhibins Alpha-inhibin gene knockout Mice develop normally at first [FSH] = 3x normal levels Mice then develop gonadal tumors Alpha-inhibin is a negative regulator of gonadal cell proliferation Alpha-inhibin is a gonadal tumor-suppressor

Mouse Models to Assess Functions of Activins Activin knockouts Knocked out activin-betaB- no activin B, activin AB, inhibin B Activin-B-deficient mice are viable but have defects in eyelid development and female reproduction (perinatal lethality of their offspring). Activin-A is up-regulated in knockout mice.

Knocked out activin- A or activin-A + activin-B Activin- A-deficient mice develop but die within 24 hr of birth Mice lack whiskers and lower incisors and have palate defects Activin-A is involved in craniofacial development Activin-A + activin-B - deficient mice show the combined defects of the two individual phenotypes, but nothing extra Activin-A and -B are therefore not functionally redundant Suggests activin AB does not have a unique role in development

Activin receptor (ActRc II) knockouts Expected them to phenocopy activin knockouts Some skeletal and facial defects, but most OK into adulthood FSH levels low, reproduction poor Suggest that this receptor important in pituitary gonadotropes

Reproductive Endocrinology III Female Reproductive Endocrinology

Follicle Development

Primary follicles Long suspension in meiotic prophase (13-50 years) Differential fate o Resting o Growing The human female has approximately 30 years of reproductive life. Only 300 -400 primary follicles reach maturity within a lifetime. Atresia (apoptosis) o The number of primary follicles: 7 million between wks 20-28 2 million at birth 300,000 at puberty

Functions of the Ovarian Follicle 1. Preserves the resident oocyte 2. Matures the oocyte at the optimal time 3. Produces hormones that develop a proliferative endometrium 4. Releases the oocyte at optimal time 5. Provides the corpus luteum, leading to implantation 6. Preserves hormonal conditions for gestation until fetoplacental metabolism is adequate Cyclic Changes in Reproductive Activity Seasons are important in some species Estrus = noun; estrous = adjective Estrous cycle in non- primates Menstrual cycle in primates o Monthly uterine maturation/sloughing sequence The Menstrual Cycle average duration of 30 days (25-35 days) Four phases o Mentrsuation: lasts about 4-5 days o Follicular phase: last for about 10-16 days o Ovularoty phase: last for about 36 hours o Luteal phase: lasts for about 14 days

Ovulation During the second half of the follicular phase, one follicle becomes dominant , undergoes further growth, and become a mature oocyte. At the end of the follicular phase, a large increase in plasma levels of gonadotropins (gonadotropin surge) takes place. 16-24 hours after the gonadotropin surge, ovulation occurs. Two essential genes: o Progesterone receptor -- stimulates production of proteolytic enzymes necessary for rupture of the follicular wall. o Cox-2 (cyclooxygenase 2)-- activates the synthesis of prostaglandins that are required for the dissociation of the mature oocyte from other cells in the follicle. o Both genes are expressed in granulosa cells and are induced by the preovulatory LH surge

The Corpus Luteum Following ovulation, the collapsed follicles becomes reorganized to form the corpus luteum. Consists of luteinized granulosa cells, luteinized thecal cells, fibroblasts, and capillaries. Produces progesterone in large amount.

Should fertilization fail to occur, the corpus luteum remains functional for 13-14 days and then undergo luteolysis, resulting in a scar-like tissue (corpus albicans).

Uterine Morphological Changes

Dynamic Changes in the Ovary, Uterus, and Serum Hormones

Endocrine Interactions in the Female Reproductive Axis

Puberty in Girls Sleep-related LH release (the first endocrine manifestation of the onset of puberty) Maturation of HPG axis Gonadotropins stimulate ovary to produce estrogens Estrogens promote female secondary sexual characteristics Somatic changes (breast development, bone maturation, fat distribution) Closure of epiphyses First menstruation (about 12 years of age)

Reproductive Cyclicity Entails the orderly repetition of the menstrual cycle Begins with the first ovulation at puberty Extends on average to about 50 years of age be interrupted by pregnancy

Menopause A period of the female climacteric during which reproductive cyclicity gradually disappears Irregular menstrual cycles Shorter intervals of the menstrual cycles The result of the loss of ovarian function An exhaustion of ovarian follicles due to repeated cycles of atresia Loss of estrogen Atrophic changes of breast, uterus, and vagina Osteoporosis Plasma gonadotropin increase

Clinical Assessment of Female Reproductive Function Plasma gonadotropin levels Plasma estradiol level Occurrence of ovulation: o Plasma progesterone level o Body temperature increases

Reproductive Endocrinology IV Female Reproductive Endocrinology Fertilization

Implantation

Placentation Pregnancy Parturition