Effect of human menopausal gonadotropin and follicle regulatory protein(s) on 3 beta-hydroxysteroid dehydrogenase in human granulosa cells.
Recently, a protein fraction [follicle regulatory protein (FRP)] which inhibits FSH-induced granulosa cell aromatase activity was isolated from both human and porcine follicular fluid. In this study, the actions of FRP on 3 beta-hydroxysteroid dehydrogenase (3 beta-HSDH) activity were examined using granulosa cells obtained from hyperstimulated patients undergoing oocyte aspiration for in vitro fertilization. Granulosa cells were cultured with 0, 167, or 500 micrograms/ml FRP with or without human menopausal gonadotropin (hMG; 10 mIU/ml). After 48 h, the medium (S) was removed and stored. Cells then were mechanically lysed and centrifuged at 10,000 X g. The supernatant was further centrifuged (100,000 X g) to obtain a microsomal fraction (M) and cytosol (C). The M fraction was resuspended in medium 199 with 10(-6) M pregnenolone plus 5 microM NAD+ and incubated for 2 h to determine 3 beta-o1 dehydrogenase activity. The S, C, and M fractions were all assayed for progesterone (P) by RIA. hMG markedly increased P concentrations in the S and C fractions. The M fraction demonstrated a hMG-dependent enhancement in 3 beta-HSDH activity. However, the hMG-associated S, C, and M P levels were decreased in granulosa cells coincubated with FRP. In conclusion, ovarian steroidogenesis may be dependent on the integrated interactions of both gonadotropins and local nonsteroidal paracrine/autocrine modulators of granulosa cell function.
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006107:Granulosa Cells / Q000201:enzymology*; D006801:Humans; D007986:Luteinizing Hormone / Q000494:pharmacology; D008596:Menotropins / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000378:metabolism; D011374:Progesterone / Q000032:analysis; D011506:Proteins / Q000494:pharmacology*; D013552:Swine
D011506:Proteins; D011374:Progesterone; D008596:Menotropins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D015096:3-Hydroxysteroid Dehydrogenases