Activin-A, but not inhibin, regulates 17beta-hydroxysteroid dehydrogenase type 1 activity and expression in cultured rat granulosa cells.
17beta-Hydroxysteroid dehydrogenase type 1 (17HSD type 1) catalyzes the reduction of estrone (E(1)) to biologically more active estradiol (E(2)). In the present study, the effect of activin, inhibin, and follistatin on 17HSD activity and 17HSD type 1 expression in cultured, unluteinized rat granulosa cells was examined. Furthermore, the effects of these hormones on 17HSD type 1 expression were compared with the expression of P450 aromatase (P450arom). Rat granulosa cells were pre-incubated in serum-free media for 3 days, followed by a 2-day treatment with activin, inhibin, follistatin and 8-Br-cAMP. Activin in increasing concentrations appeared to effect a dose-dependent increase in 17HSD activity. In addition, increasing concentrations of activin also increased 17HSD type 1 mRNA expression. Addition of 8-Br-cAMP at concentrations of 0.25 and 1.5 mmol/l together with activin significantly augmented the stimulatory effects of activin alone in the cultured cells. Neither inhibin, nor follistatin, either alone or in combination with 8-Br-cAMP, had any notable effects on 17HSD activity and 17HSD type 1 expression. Preincubation of activin with increasing concentrations of follistatin significantly diminished the stimulatory effect of activin. In the presence of follistatin, activin did not significantly increase the 8-Br-cAMP-induced 17HSD activity and 17HSD type 1 expression. The culturing of granulosa cells in the presence or the absence of inhibin or follistatin with or without 8-Br-cAMP did not alter the effect of these peptides on P450arom expression in rat granulosa cells as judged by Northern blot analysis of total RNA. However, cAMP-induced P450arom expression was enhanced by activin treatment, except when follistatin was present. This is in line with the suggested role of follistatin as an activin-binding protein, which limits the bioavailability of activin to its membrane receptors. Thus, the results support the notion of a paracrine/autocrine role of activin in follicular steroidogenesis of growing follicles.
Ghersevich
S
S
Biocenter Oulu and WHO Collaborating Center for Research on Reproductive Health, University of Oulu, PO Box 5000, Kajannintie 50, FIN-90401, Oulu, Finland.
Akinola
L
L
Kaminski
T
T
Poutanen
M
M
Isomaa
V
V
Vihko
R
R
Vihko
P
P
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D028341:Activins; D000818:Animals; D001141:Aromatase / Q000235:genetics; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004960:Estradiol Dehydrogenases / Q000235:genetics*; D005260:Female; D038681:Follistatin; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006023:Glycoproteins / Q000494:pharmacology; D006107:Granulosa Cells / Q000201:enzymology*; D006801:Humans; D007265:Inhibins / Q000494:pharmacology*; D007700:Kinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D038681:Follistatin; D006023:Glycoproteins; D011994:Recombinant Proteins; D028341:Activins; D015124:8-Bromo Cyclic Adenosine Monophosphate; D007265:Inhibins; D004960:Estradiol Dehydrogenases; D001141:Aromatase