Pathophysiology of the Reproductive Cycle Molecular medicine case studies : Mutations of the FSH and LH receptors (which have similar structure) Prima

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1 FEMALE REPRODUCTIVE HORMONE BIOCHEMISTRY DEPARTMENT

2 Pathophysiology of the Reproductive Cycle Molecular medicine case studies : Mutations of the FSH and LH receptors (which have similar structure) Primary ovarian failure caused by a point mutation in the FSH receptor gene. Chromosome 2p C to T in nucleotide 566 of exon 7 of the FSH receptor gene. Alanine to Valine substitution at residue 189 of the protein.

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4 Pituitary Hormones Posterior pituitary Stores and releases 2 hormones that are produced in the hypothalamus: o Antidiuretic hormone (ADH/vasopressin): o Oxytocin:

5 Oxytocin 9 amino acid peptide, produced d primarily il by paraventricular nucleus and small amount by supraoptic nucleus Targets: Uterus and mammary glands Contraction ti of pregnant uterus Stimulates contractions of the mammary gland alveoli. li Milk-ejection reflex.

6 Anterior pituitary Pituitary Hormones Prolactin Growth Hormone (GH) Thyroid Stimulating Hormone (TSH) Adrenocorticotropic t i Hormone (ACTH) Follicle-stimulating Hormone (FSH) Luteinizing i i Hormone (LH)

7 PROLACTIN Protein hormone of 199 amino acids. In females it stimulates milk production by the mammary glands.. Release is inhibited in non-pregnant women. As estrogen and progesterone levels rise late in pregnancy, it stimulates prolactin release. Hyperprolactinaemia can cause menstrual problems in females and breast enlargement in males.

8 Hypo-Pit Pit-Gonadal Axes

9 HORMONAL CONTROL OVER GONADAL FUNCTION An Overview GnRH Lh+fsh estrogens androgens Hypothalamus Anterior pituitary Gonads - (synthesizes Gonadotropin (synthesizes Gonadotropins: synthesize gonadal hormones releasing luteinizing hormone hormone) follicle stimulating hormone)

10 RESEPTOR Reseptor Hormon umumnya di membran plasma atau didalam sitoplasma. Interaksi H-R signal transduction kedalam sel. Reseptor mengalami up and down Regulation TRH,TSH TSH dan T3-T4 T4 : regulasi melalui reseptor masing-masing dalam mekanisme negative feedback.

11 RESEPTOR Hal ini dipengaruhi oleh a.l. 1. jumlah hormon tsb yang disekresikan, misalnya reseptor Insulin akan up regulate bila Insulin sedikit 2. pengaruh dari hormon lain,misalnya reseptor LH akan meningkat oleh sekresi FSH Struktur reseptor LH mirip dengan reseptor FSH, hcg sel luteal dan TSH Reseptor-reseptor reseptor tersebut dapat bermutasi timbul kelainan.

12 RESEPTOR Beberapa obat dapat berinteraksi dengan reseptor tersebut Efek yang timbul : agonist (menyerupai efek ligand yang sebenarnya) atau antagonist (berlawanan dengan efek ligand nya, misalnya menghambat transduksi signal) Reseptor juga dipunyai oleh banyak molekul lain, seperti Interleukin, Interferon, Antigen, Neurotransmiter, LDL dll.

13 Action of GnRH A system stimulation of phosphatidylinositol is GnRH action GnRH binds to receptors in the cell membrane of the gonadotrope Phospholipase C is activated, catalyzes the hidrolysis of PIP 2 to form DAG and IP 3 DAG activates protein kinase C (PKC), which phosphorylates specific protein It participate in the resulting secretory process to transport LH and FSH to cell exterior IP 3 stimulates of Ca 2+ from calcium stroge particle, cause increased stimulation of PKC participates in exocytosis of LH and FSH from cell

14 Action of GnRH

15 GONADOTROPINS Follicle-stimulating hormone (FSH) Target: Ovaries and Testes Stimulates the maturation of sperm and egg, in the females causes the release of estrogen

16 GONADOTROPINS Luteinizing Hormone (LH) Target: Ovaries and Testes Male: Production of testosterone In females works with FSH to cause follicle development, and then independently is responsible for ovulation.

17 Sex Hormones Gonad and Adrenal Estrogen Progesterone Dihydrotestosterone Testosterone

18 Ovarian hormones Steroids o Estrogens o Androgens o Progesterone Peptides

19 A Steroid Molecule

20 OVARIAN STEROID HORMONES Produced in both interstitial and follicular l cells Derivatives of cholesterol (coming from LDL-lipoproteins and de novo synthesis) s)

21 Stimulasi sintesis hormon steroid

22 SYNTHESIS

23 ESTROGEN o Chemical structure: C18 Source: follicular cells o corpus luteum o placenta o adrenal cortex o adipose tissue (DHEA androstenedione E1)

24 SYNTHESIS First step happens in mitochondria All the rest in smooth ER Reduction in number of carbon atoms Estrogens are primarily synthesized in granulosa cells Aromatization of Androstenedione and Testosterone within tissue is a major source of Estrogens in the male and post menopausal female.

25 Ovarian Steroid Hormones Estrogen produced by thecal cells : the major steroid in the circulation Estrogen synthesized by granulosa cells : a local role, possibly related to ovum maturation FSH regulate production and subsequent induction and activation of Aromatase in the granulosa cells Estradiol (E2) is oxidized to Estrone within the liver and Estrone hydrated to Estriol (E3).

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27 Estrogens in circulation Estradiol o Secreted by the ovary Estrone o Derived from peripheral conversion of estradiol and androstenodione Estriol o Liver metabolite of estrone

28 TRANSPORT IN BLOOD Mostly carrier-bound (albumin, sex hormone-binding globulin) Two peaks o Before ovulation o Mid-luteal peak Levels : much higher in the follicular fluid than in the plasma!

29 Mechanisms of Action Cytoplasmic receptor binds to steroid hormone. Translocates to nucleus. DNA-binding domain binds to specific HRE of the DNA. Dimerization occurs. Process of 2 receptor units coming together at the 2 half-sites. Stimulates transcription of particular genes.

30 Mechanisms of Action

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32 Metabolism Metabolized to glucuronides and sulfates Recycled in enterohepatic circulation Excreted in urine

33 Physiological Development and maintenance of uterus, uterine tubes,vagina, external genitalia and breasts Cyclic changes in the endometrium, cervix, vagina Growth of the ovarian follicles Motility of the uterine tubes

34 Physiological Pregnancy: uterine muscle mass, excitability, breasts Female secondary sex characteristics (fat deposits, etc) Estrous behavior in animals, increased libido in humans

35 Progesteron The most distinctive hormone between males and females Chemical structure: C21 Source: o c. luteum o placenta o follicles (small amount) o adrenal cortex

36 BIOSYNTHESIS : FSH promotes the growth of the follicle by acting through receptors on the granulosa cells and inducing i the Aromatase enzyme to convert Androgens to Estrogens. The Androgens originate from the theca which is stimulated by LH through its receptors initially i i on theca cells and appear in the granulosa cells and are probably coupled to the same second messenger (camp) as FSH receptors

37 BIOSYNTHESIS Progesterone synthesis is an early step in the biosynthesis of androgen and estrogen within the thecal cell. But circulating progesterone is produced by corpus luteum

38 Transport 2% free 80% albumin-boundbound 18% corticosteroid-binding protein (transcortin) -bound

39 Physiological Cyclic changes in the endometrium, cervix, and vagina Breasts: supports the secretory function during lactation Inhibits LH secretion Responsible for preparing the reproductive tract for implantation and the maintenance of pregnancy

40 Mechanism of action of Ovarian Steroid Hormones E2 and Progesterone interact t with cytoplasmic and/or nuclear protein receptors. Release of the two receptors subunits with attached steroid hormones from association with a heat-shock protein. Either identical subunits un singly or possibly together interact directly with the DNA hormone responsive element to activate transcriptional events leading to translation of a cell-specific protein.

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42 Androgens Androstenedione and Testosterone as a precursors of estrogens are secreted by ovaries in the beginning of Puberty. Development of secondary sexual characters almost totally by the actions of estrogens.

43 Androgens Androgens of ovarian origin may be influenced at a puberty. Acne, results from sebaceous gland activation by ovarian androgens. Pubic and axillary hair growth are also attributed to androgens of ovarian and/or adrenal origin. Changes in enzymic (3β-hydroxysteroid dehydrogenase, 1-hydroxylase and 17,20-desmolase) activity, increased production of adrenal androgens

44 OVARIAN PEPTIDE HORMONES Relaxin o Relaxes pelvic joints o Softens and dilates cervix o Sperm mobility - in males Inhibin o Selective inhibitory control of FSH Activin o Selective stimulaton of FSH o Cell differentiation

45 OVARIAN PEPTIDE HORMONES Follistatins o Inhibit FSH secretion Gonadotropin surge attenuating factor o Prevents premature LH surge

46 Intrafollicular Polypeptide Regulatory Factors Several peptides have been isolated from ovarian follicles and regulate oocyte maturation : 1. Oocyte Maturation ti Inhibitor 2. Ovarian Growth Factors 3. Inhibins 4. Activins

47 Intrafollicular Polypeptide Regulatory Factors 1. Oocyte Maturation Inhibitor : Low molecular weight-peptide Its concentration in antral fluid decreases as the follicle maturates Gap junctions, present between cells of the corona radiata and the oocyte structural pathway for cell-to to-cell communication. OMI may exert its action indirectly through an action on cumulus cells LH induced oocyte maturation may result from an uncoupling between the cumulus cells and the oocyte

48 Intrafollicular Polypeptide Regulatory Factors 2. Ovarian Growth Factors : Induction of LH receptors is a critical aspect of granulosa cell differentiation and ovarian follicular development Pituitary FSH functions as the inducer of granulosa cell LH receptor acquisition This FSH-dependent LH receptor induction is inhibited by EGF and FGF

49 Intrafollicular Polypeptide Regulatory Factors 3. Inhibin : Inhibin is present in follicular fluid, ovarian and granulosa cell extracts Inhibin exerts a suppressive action on both basal and GnRH-stimulated pituitary FSH secretion Circulating levels of Inhibin are inversely related to plasma FSH levels. Inhibin can be detected in the plasma of women given FSH, but not in castrated subjects.

50 Intrafollicular Polypeptide Regulatory Factors 3. Inhibin : Inhibin composed of two subunits that t are linked by disulfide bridges The two subunits (α, β) ) encoded d by separated genes The control of inhibin biosynthesis is at the level of transcription.

51 Intrafollicular Polypeptide Regulatory Factors 4. Activin : Purified fractions of follicular fluid exhibited stimuli rather than inhibitory effects on FSH release from pituitaries invitro. β Subunits of inhibin and transforming growth factor β (TGFβ) were similar. Dimers of the β subunits of inhibin, β A β B, β A β A and β B β B were named Activin, Activin A and Activin B Activin can enhance basal secretion of FSH in a dose-dependent dependent manner in vitro without affecting the secretion of LH.

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