Follicle-Stimulating Hormone Regulation of Cytochrome P-450 Side-Chain Cleavage Messenger Ribonucleic Acid Accumulation by Porcine Granulosa Cells Isolated from Small and Medium Follicles1

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Abstract
The experiments described here were conducted to examine regulation of cytochrome P-450 side-chain cleavage (SCC) mRNA accumulation in procine granulosa cells isolated from small (1-4 mm) and medium (5-6 mm) follicles. Granulosa cells were cultured under the following conditions: 1) for 48 h or 96 h with 0, 50, or 200 ng/ml porcine FHS; 2) for 96 h with 200 ng/ml FSH and aminoglutethimide (100 .mu.M); and 3) for 96 h with forskolin (100 .mu.M). Total RNA was extracted and examined by Northern and dot-blot hybridization analysis, and culture media were assayed for progesterone concentration. Northern blot analysis revealed a single band approximately 2.1 kb in size. Accumulation of SCC mRNA by granulosa cell was both FSH dose- and culture dependent (p < 0.05) with maximal increases approximately 4.5 times control levels. Aminoglutethimide reduced progesterone production by about 80% while having no effect on granulosa cell accumulation of SCC mRNA compared to cells stimulated with 200 ng/mol of FSH. Forskolin-treated cells produced significantly more progesterone than did cells treated with FSH, but accumulation of SCC mRNA was similar. In respnose to FSH, concentration of SCc mRNA did not vary with follicle size, but granulosa cells from small follicles produced significantly more progesterone than did those from medium follicles. These results demonstrate that concentration of SCC mRNA in cultured porcine granulosa cells is FSH dose-dependent, does not vary significantly in cells from small- and medium-sized follicies, and is correlated with progesterone production, but may not parallel progesterone secretion. This last observation indiates that control at sites other than SCC mRNA can affect progesterone production.

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