REGULATION OF INHIBIN ACTIVIN SUNBUNIT MESSENGER RIBONUCLEIC-ACIDS (MESSENGER-RNAS) BY ACTIVIN-A AND EXPRESSION OF ACTIVIN RECEPTOR MESSENGER-RNAS IN CULTURED HUMAN GRANULOSA-LUTEAL CELLS

Recent studies have indicated that activin and inhibin may act as local regulators of cell growth and steroidogenesis in the human ovary. We studied the effect of recombinant human activin A and purified bovine inhibin A on the steady state messenger RNA (mRNA) levels of the inhibin/activin alpha-, beta(A)-, and beta(B)-subunits in cultured granulosa-luteal (GL) cells from preovulatory ovarian follicles of women undergoing in vitro fertilization. Activin A induced the expression of a 4.8-kilobase beta(B)-subunit mRNA transcript without affecting basal expression levels of the alpha- and beta(A)-subunit mRNAs. It stimulated beta(B)-subunit mRNA levels in a concentration- and time-dependent manner. Maximal stimulation of beta(B)-subunit mRNA levels was obtained with 30-100 ng/ml activin A. The level of beta(B)-subunit mRNAs increased significantly 8 h after stimulation, rising gradually thereafter to a maximum at 48 h. Inhibin A did not affect the mRNA levels of any inhibin/activin subunits, nor did it inhibit the effect of activin A. Recombinant human follistatin did not affect basal beta(B)-subunit mRNA levels, but it neutralized the effect of activin A. Although hCG induces inhibin/activin alpha- and beta(A)-subunit mRNA levels in human GL cells, it did not increase basal beta(B)-subunit levels. By contrast, it inhibited activin A-induced beta(B)-subunit mRNA levels. On the other hand, activin A decreased hCG-induced mRNA levels of the inhibin alpha-subunit and cytochrome P450 side-chain cleavage (P450scc) enzyme, an important rate-limiting enzyme in human GL cell progestin synthesis. Moreover, we observed by Northern blot analysis that cultured human GL cells as well as freshly isolated preovulatory granulosa cells express the specific mRNAs for all currently known serine/threonine kinase activin receptors, i.e. activin receptors I, IB, II, and IIB. Our results suggest that in GL cells, activin A may locally stimulate synthesis of the beta(B)-subunit in an autocrine or paracrine manner, and that in human ovary, regulation of the beta(B)-subunit differs from that of the alpha- and beta(A)-subunits.