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Gonadotropin-Induced 14-Reductase and 7-Reductase Gene Expression in Cumulus Cells during Meiotic Resumption of Porcine Oocytes

Department of Applied Animal Science (Y.Y., M.N., T.T., M.S.), Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan; and Department of Technology and Resources (N.I.), Graduate School of International Development and Cooperation, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8529, Japan

Address all correspondence and requests for reprints to: Masayuki Shimada, Ph.D., Department of Applied Animal Science, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan. E-mail: mashimad{at}hiroshima-u.ac.jp.

Progesterone is produced from cholesterol in cumulus cells during meiotic resumption of porcine oocytes. In follicular cells, it has been shown that exogenous lipoprotein-bound cholesterol ester can be used for steroid hormone production. However, in serum-free medium, progesterone is also secreted by FSH- and LH-stimulated cumulus-oocyte complexes, suggesting that progesterone could be produced from de novo synthesized cholesterol in cumulus cells. In the present study, we investigated the expression of 14-reductase and 7-reductase, which are the members of the superfamily that converts acetyl-CoA to cholesterol in cumulus cells. The expression of both genes was analyzed by RT-PCR. Both 14-reductase mRNA and 7-reductase mRNA in cumulus cells, cultured until 4 h, were under the level of detection limit. In response to gonadotropins, both mRNA levels were dramatically up-regulated, reaching a maximum at 20 h. To clarify the role of induced enzymes in cumulus cells, cumulus-oocyte complexes were cultured with either 14-reductase inhibitor, AY9944-A-7, or 7-reductase inhibitor, BM15.766. The results indicated that these inhibitors significantly suppressed the progesterone production in cumulus cells and meiotic progression of oocytes. The inhibitory effects reached a maximum at 1 µM AY9944-A-7 or 20 µM BM15.766. The addition of 20 ng/ml progesterone overcame the inhibitory effects of both drugs on meiotic resumption of oocytes. These results imply that gonadotropin-induced expression and function of 14-reductase and 7-reductase in cumulus cells contribute to oocyte meiotic resumption via a progesterone-dependent pathway.

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