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Effect of Follicle-Stimulating Hormone on Steroid Secretion and Messenger Ribonucleic Acids Encoding Cytochromes P450 Aromatase and Cholesterol Side-Chain Cleavage in Bovine Granulosa Cells In Vitro¹

^a Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada J2S 7C6

We determined 1) whether the previously observed induction of estradiol secretion in bovine granulosa cells cultured in serum-free conditions is associated with an increase in cytochrome P450 aromatase (P450_arom) mRNA abundance and 2) whether P450_arom mRNA levels are responsive to FSH in vitro. Granulosa cells from small (2–4-mm) follicles were cultured in serum-free medium. Estradiol secretion increased with time in culture and was correlated with increased P450_arom mRNA abundance. Progesterone secretion also increased with time in culture, but P450 cholesterol side-chain cleavage (P450_scc) mRNA abundance did not.

FSH stimulated estradiol secretion and P450_arom mRNA abundance; the effect was quadratic for both estradiol and P450_arom mRNA. Estradiol secretion and P450_arom mRNA levels were correlated. FSH stimulated progesterone secretion and P450_scc mRNA abundance, although the minimum effective dose of FSH was lower for estradiol (0.1 ng/ml) than for progesterone (10 ng/ml) production. Insulin alone stimulated estradiol secretion and P450_arom mRNA levels but not progesterone or P450_scc mRNA abundance. We conclude that this cell culture system maintained both estradiol secretion and P450_arom mRNA abundance responsiveness to FSH and insulin, whereas P450_scc mRNA abundance and progesterone secretion were responsive to FSH but not insulin.

First decision: 24 June 1999.

¹ This work was supported by grants from NSERC Canada and CONACYT, Mexico.

² Correspondence: C.A. Price, CRRA, Faculté de médecine vétérinaire, Université de Montréal, 3200 Sicotte, St-Hyacinthe, PQ, Canada J2S 7C6. FAX: 450 778 8103; pricec{at}medvet.umontreal.ca

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