Steroid Hormones

Steroid hormones
Glucocorticoids. Mineralocorticoids. Sex steroids: -Progesterone. -Androgens. -Estrogens.

Steroid hormones
Pregnane (C21): Progestins and
corticoids.

Androstane(C19): Androgens.
Estrane (C18): Estrogens

Cholesterol

Steps:
Acquisition of cholesterol. Transfer of cholesterol from outer to inner mitochondrial membrane. Side chain cleavage to pregnenolone. 4 pathway (progesterone) or 5 pathway (17 hydroxy pregnenolone) to androstenedione. Aromatization to corresponding estrogen.

Steps:
I : Acquisition of cholesterol:
From blood through LDL.

De-novo synthesis.

Cholesterol Metabolism: Uptake

Steroid producing organs

Steps:
II: Transfer of cholesterol from outer to inner mitochondrial membrane: Rate limiting step. Sensitive to trophic hormones. Involve carrier proteins StAR, SCP2, SAP, PBR. Deficiency of StAR causes congenital lipoid adrenal hyperplasia

Mitochondria

Mitochondria

Steps:
III: Cholesterol side chain cleavage: Achieved by hydroxylation at 20 and 22 position then cleavage. Carried out by single mitochondrial CYP450 named CYP450scc. Produce pregnenolone which is the mother molecule for all steroids

Steroidogenic enzymes:
Cytochrome P 450(CYP 450). Family of oxidases. Contain Haem group for oxygen carriage. Mitochondrial or microsomal. Utilizes NADH as coenzyme. Dehydrogenases 3 hydroxy steroid dehydrogenase. 17 hydroxy steroid dehydrogenase.

Cellular Locations of Steroid Synthesis

sER

Smooth Endoplasmic reticulum (sER)

sER

Orphan receptors and steroidogenesis:

Transcription factors that belong to nuclear receptors superfamily. Yet unidentified ligands. Include steroidogenic factor-1 and DAX-1. Steroidogenic factor-1 influences expression of StAR, Steroidogenic enzymes and others.

Ovarian Steroidogenesis
Sites: Maturing follicles and CL Maturing follicles : 2 adjacent important populations:
CL important cells derived from follicle after ovulation
Granulosa lutein Theca lutein Granulosa cells Theca interna cells

Granulosa cells lack P450scc

No cholesterol pregnenolone Lack P450c17 No further androgen synthesis Rely on theca cell androgens Insensitive to LH signal for androgen production.
Just prior to ovulation. Small numbers LH receptors expressed. Important to luteinization.

EXCEPT when advanced follicle developed

Granulosa cells Sensitive to FSH

FSH stimulation
Proliferation granulosa cells Secretion inhibin B Expression of aromatase.

Granulosa cells express aromatase

Androgens estrogens Most important source of E2

Figure 9. The early differentiation of the granulosa cells during preantral folliculogenesis involves the expression of FSH receptors. Animal studies support the concept that this process involves an activin autocrine/paracrine mechanism. (Erickson GF: Dissociation of Endocrine and Gametogenic Ovarian Function. In Lobo, R. (ed.): Perimenopause. Serono Symposia, Springer-Verlaag, 1997. Reproduced with permission from Springer-Verlag, New York.)

Theca (interna & externa) Express P450scc

Cholesterol pregnenolone Sensitive to LH
cAMP PKA StAR PBR P450scc activity in inner mitochondrial membrane

Pregnenolone 5 (androgen) pathway

Uses P450c17

Preovulatory LH surge
Ovulation Luteinization theca interna, granulosa Development CL

 Theca interna express aromatase (androgens estrogens)

BUT much less active than granulosa. Once ovulation occurs, expression ceases. So decreased secretion of estrogens from CL

Follicular 2-Cell Pathway

The 2 cell 2 gonadotropin theory

Granulosa lutein cells express P450scc

So can produce pregnenolone

BUT granulosa lutein cells lack P450c17

No androgen synthesis from pregnenolone
Rely on theca lutein cells for androgens

BUT progesterone synthesis from pregnenolone ( 4 pathway). Insensitive to LH signal to produce androgens
BUT small numbers LH receptors expressed in advanced follicle stimulates secretion of progesterone, and inhibin A

Coupling of steroidogenesis to Gametogenesis

Ovarian follicles named by maturation stage Primordial
Primary oocyte Squamous epithelium; single layer
Growth & remodeling

40 m; oocyte 15-25 m Quiescent

 Primary follicle Oocyte enlarges (80-100 m) Follicle epithelium simple cuboidal Zona pellucida (glycoprotein layer) deposits Stroma begins to organize as theca

Secondary=preantral follicle

Granulosa proliferates, stratifies

Signaled by oocyte growth factors
Growth Differentiation Factor 9 (GDF-9) Bone Morphological Protein 15 (BMP-15)

Granulosa cells produce activin Granulosa cells become sensitive to FSH

Important to selection Important to differentiation Important to maturation

Mitosis and rearrangement result in Diff layers have diff morphology & function

Secondary=preantral follicle
Final 2o follicle 100-200 m
Fully grown oocyte with zp Approx 9 layers granulosa Basal lamina Theca interna with capillary network Theca externa

Secondary=preantral follicle Granulosa cells connected by gap junctions

Synchronization Nutrient transfer

Basement membrane develops When 6 layers granulosa

Definitive theca layer Angiogenesis blood supply to theca

Granulosa cells

Theca cells

Tertiary=antral follicle
Chamber Follicular fluid contains hormones
Blood-derived From oocyte From granulosa cells

Cavitation signaled by follicle cells Granulosa cells produce inhibin B and aromatize androgens. Theca fully developed

Tertiary=antral follicle

Theca interna = steroid producing (androstenedione)

Theca externa

Extensive sER, cytoplasmic lipid droplets, tubular mitochondrial cristae Receptors for LH, insulin Richly vascularized

Smooth muscle cells Autonomic neurons

Figure 12. Photomicrograph of an early tertiary follicle 0.4 mm in diameter at the cavitation or early antrum stage. zona pellucida (ZP); granulosa cells (GC); basal lamina (BL); theca interna (TI); theca externa (TE); granulosa mitosis (arrowheads). (Bloom W, Fawcett DW In A Textbook of Histology. Philadelphia, WB Saunders Company, Philadelphia 1975. With permission from Arnold.)

Preovulatory=Graafian follicle
Now single, dominant follicle
Overall size varies; determined by amount of follicular fluid. Granulosa differentiated into mural, antral , cumulus. Produce 400 estradiol/day. LH surge induces completion 1st meiotic division 2o oocyte
2nd meiotic division @ fertilization

Ovulation
Induced by LH surge Rupture
Follicle wall Follicle cell layers

Oocyte with corona radiata expelled in ooze like manner.

Luteinization
Initiated by LH surge CL forms from
Granulosa lutein. Theca lutein. Associated microvasculature invade granulosa. If no fertilization, involution & apoptosis 14 d after formation.

Defined life span

Steroid hormone receptors

Nuclear receptors. Dimerize and bind to DNA at specific sites called steroid response elements. Binding of ligand bound receptor to DNA stimulates transcription of specific proteins to mediate response.

Conclusion:
Cholesterol is the basic building block of all steroids. Cholesterol can be synthesized by all steroid producing organs except placenta but local production is insufficient so mostly obtained from blood LDL.

Conclusion:
Steroidogenesis occur in 2 cellular sites; mitochondria and sER. Steroidogenic enzymes belong to 2 families CYP 450 and dehydrogenases. After pregnenolone 2 pathways according to site.

Conclusion:
Ovarian steroidogenesis depends on cooperation between theca cells under the effect of LH and granulosa cells under the effect of FSH (the 2 cell 2 gonadotropin theory. Ovarian steroidogenesis unlike testicularis coupled to gametogenesis.

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