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Experiments were designed to test the hypotheses that (1) follistatin gene expression in granulosa cells is regulated during follicular growth, and (2) that alteration of follistatin mRNA concentration can be hormonally induced in primary cultures of porcine granulosa cells. RNA isolated from granulosa cells from small (1–3 mm diameter), medium (3–5 mm) and large (> 5 mm) follicles of prepubertal and postpubertal sows was analysed by hybridization to a porcine follistatin cDNA probe. Amounts of follistatin mRNA increased with follicular diameter, but no differences in follicular follistatin mRNA were detected between prepubertal and postpubertal sows. Treatment of cultured porcine granulosa cells with FSH or LH for 20 h stimulated follistatin mRNA concentration by a factor of two (100 ng FSH ml−1) and a factor of 1.5 (10 ng LH ml−1), respectively, over untreated controls. Treatment of cultured granulosa cells with 200 ng FSH ml−1, 200 ng LH ml−1, 10 μmol dibutyryl cAMP l−1, 30 μmol forskolin l−1 and 100 ng cholera toxin ml−1 stimulated follistatin mRNA accumulation in granulosa cells by factors of 4.9, 3.7, 1.6, 13.7 and 3.5, respectively, compared with control cultures. Stimulation of follistatin mRNA accumulation in cultured granulosa cells by dibutyryl cAMP (30, 100 and 300 μmol l−1) and forskolin (3, 10 and 100 μmol l−1) was dose dependent. FSH and forskolin induced time-dependent increases in follistatin mRNA concentration in cultured granulosa cells, with maximal induction occurring 72 h after treatment (a factor of 4.5 for FSH and 15.5 for forskolin). These results demonstrate that (1) increased follistatin mRNA in granulosa cells is associated with increased follicular diameter, (2) increased follistatin mRNA in cultured porcine granulosa cells can be induced by gonadotrophins, and (3) the gonadotrophin-induced increase in follistatin mRNA accumulation in cultured granulosa cells can be mimicked by agents that increase intracellular cAMP concentrations.
