Molecular Human Reproduction, Vol. 8, No. 11, 992-997,
November 2002
© 2002 European Society of Human Reproduction and Embryology
Ovary and oogenesis |
Regulation of follistatin-related gene (FLRG) expression by protein kinase C and prostaglandin E2 in cultured granulosa-luteal cells
1 Department of Pathology, P.O. Box 21, University of Helsinki, FIN-00014 Helsinki, 2 Department of Paediatrics, P.O. Box 1777, Kuopio University and University Hospital, FIN-70211 Kuopio, and 3 Department of Obstetrics and Gynaecology, P.O. Box 140, Helsinki University Central Hospital, FIN-00290 Helsinki, Finland
Activin and its binding protein follistatin may act as local regulators of cell growth and steroidogenesis in the human ovary. The recently identified follistatin-related gene (FLRG) is expressed abundantly in the human ovary, has high affinity for activin, and is able to inhibit activin-induced transcriptional responses. However, little is known about the regulation of FLRG expression in specific cell types in the ovary, while it is known that gonadotrophins induce follistatin gene expression in human granulosa-luteal cells. In this study, we investigated the expression of FLRG mRNA in granulosa-luteal cells of preovulatory follicles obtained from women undergoing IVF. FLRG mRNA was detected by RT–PCR in fresh and cultured granulosa-luteal cells, as well as in normal ovarian stroma, theca and granulosa cells. Northern blot analysis revealed a 2.5 kb transcript of the FLRG in cultured granulosa-luteal cells. The protein kinase C activator, 12-O-tetradecanoyl phorbol 13-acetate (TPA, 160 nmol/l), and prostaglandin E2 (PGE2, 1 µmol/l) increased FLRG mRNA accumulation up to 3–8 fold over the control level after 24 h of treatment, and these stimulatory effects were dose-dependent. Co-treatment with the protein kinase C inhibitor, Ro-31–8220 (3 µmol/l), blocked the stimulatory effect of TPA. Although short term treatment with the protein kinase A activator, (Bu)2cAMP (1 mmol/l), slightly reduced FLRG mRNA expression in most experiments, long term treatment with FSH (100 IU/l), LH (100 IU/l), or (Bu)2cAMP had no significant effect on the FLRG mRNA levels. As expected, gonadotrophins, protein kinase A and C activators and PGE2 increased granulosa-luteal cell progesterone secretion into the culture media. Taken together, previous and our present data suggest that protein kinase C and A signal transduction pathways differently regulate the expression of FLRG and follistatin genes in human ovarian granulosa-luteal cells.
follistatin-related gene/gonadotrophin/granulosa-luteal cells/protein kinase A/protein kinase C
4 To whom correspondence should be addressed. E-mail: Jiangi.Liu{at}helsinki.fi
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