Abstract:
:FSH stimulates follicular growth through inducing granulosa cell proliferation. Our working hypothesis is that mitogenesis is facilitated by a locally produced growth/differentiation factor(s) that modulates FSH action in developing granulosa cells. The present study was undertaken to examine the effect of the related gonadal peptides activin and inhibin on granulosa cell proliferation. Monolayer cultures of granulosa cells isolated from preantral/early antral follicles in immature rat ovaries were established by incubation overnight in serum-free medium 199. Treatment was initiated with serum-free medium containing recombinant human (rh) activin and/or rh-inhibin in the presence or absence of FSH. After incubation for 18 h, medium was collected for progesterone determination (as a marker of cell differentiation), and the cell monolayers were incubated for 2 h in the presence of [3H]thymidine to measure DNA synthesis. Activin dose dependently (1-100 ng/ml) stimulated DNA synthesis (minimal effective dose, 1 ng/ml), whereas inhibin or FSH alone was without effect. When activin (1 ng/ml), but not inhibin, was present with FSH, the gonadotropin caused dose-dependent increases in [3H]DNA synthesis over a wide range of FSH concentrations (1-100 ng/ml). This property of activin was unaltered by the additional presence of inhibin (1-100 ng/ml). To analyze the role of cAMP in mediating the mitogenic action of FSH in the presence of activin, experiments were repeated substituting a membrane-permeable cAMP agonist, 8-bromo-cAMP (8br-cAMP; 0.1-3 mM). Similar to FSH, 8br-cAMP had no effect on granulosa cell DNA synthesis in the absence of activin. However, in the presence of activin (1 ng/ml) 8br-cAMP was stimulatory. The dose response to 8br-cAMP revealed a biphasic effect on DNA synthesis and differentiation: DNA synthesis rose to a maximum in the presence of 0.5 mM 8br-cAMP and declined thereafter. Progesterone synthesis only started to increase in the presence of 0.1 mM 8br-cAMP, rising to a maximum at 3 mM 8br-cAMP, at which concentration DNA synthesis was fully suppressed. We conclude that activin induces DNA replication in rat granulosa cells. In the presence of activin, FSH and 8br-cAMP are mitogens. These actions of FSH and 8br-cAMP occur at doses too low to stimulate steroidogenesis, emphasizing the role of intracellular cAMP tone in granulosa cell proliferation and differentiation.
journal_name
Endocrinologyjournal_title
Endocrinologyauthors
Miró F,Hillier SGdoi
10.1210/endo.137.2.8593790subject
Has Abstractpub_date
1996-02-01 00:00:00pages
464-8issue
2eissn
0013-7227issn
1945-7170journal_volume
137pub_type
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