Transcriptional and post-transcriptional regulation of FSH receptor in rat granulosa cells by cyclic AMP and activin.
The effect of FSH on the induction of FSH receptors in granulosa cells is believed to be mediated, at least in part, by the cAMP second messenger system. We examined the effect of activin and cAMP on FSH receptor expression in this culture system. Steady-state levels of FSH receptor mRNA, analyzed by Northern blot hybridization, increased 3.5-fold in response to 24-h incubation with activin and 1.7-fold with 12-h incubation with 8-bromoadenosine 3,5-cyclic monophosphate (8-Br-cAMP; 0.2 mM). We have investigated whether 8-Br-cAMP- and/or activin-induced increases in FSH receptor mRNA levels are the result of increased transcription and/or altered mRNA stability. The rates of FSH receptor mRNA gene transcription, assessed by nuclear run-on transcription assay, increased 3-fold in cells treated with activin and 1.5-fold in cells treated with 8-Br-cAMP for 2 h. To examine the degradation rates of FSH receptor mRNA transcripts, granulosa cells were preincubated with 8-Br-cAMP, activin, or medium alone for 6 h. After the preincubation period, 5 microM actinomycin-D or 200 microM 5,6-dichloro-1-beta-ribofuranosyl benzimidazole were added to arrest new RNA synthesis. The decay curves for the 2.4 kb FSH receptor mRNA transcript in granulosa cells were not significantly different in the absence or presence of 8-Br-cAMP. Activin, on the other hand, significantly altered the slope of the FSH receptor mRNA decay curve and increased the half-life of the 2.4 kb FSH receptor mRNA transcript. These data provide evidence that cAMP induces FSH receptor mRNA levels by stimulating the transcription rate and that activin increases FSH receptor mRNA levels both by stimulating transcription rates and by stabilizing the FSH receptor mRNA transcripts.
Tano
M
M
Department of Obstetrics and Gynecology, Gunma University School of Medicine, Japan.
Minegishi
T
T
Nakamura
K
K
Karino
S
S
Ibuki
Y
Y
Miyamoto
K
K
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D028341:Activins; D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology*; D007265:Inhibins / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D011962:Receptors, FSH / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D013268:Stimulation, Chemical; D014158:Transcription, Genetic / Q000187:drug effects*
D006133:Growth Substances; D012333:RNA, Messenger; D011962:Receptors, FSH; D028341:Activins; D015124:8-Bromo Cyclic Adenosine Monophosphate; D007265:Inhibins; D000242:Cyclic AMP