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Human Reproduction, Vol. 11, No. 3, pp. 497-502, 1996
© 1996 European Society of Human Reproduction and Embryology

An aminopeptidase inhibitor, bestatin, enhances progesterone and oestradiol secretion by porcine granulosa cells stimulated with follicle stimulating hormone in vitro

Toshiyuki Tachibana1, Hiroshi Fujiwara1,4, Hiroshi Suginami3, Kimihiko Nakamura1, Tetsuro Honda1, Shigetoshi Yamada1, Michiyuki Maeda2 and Takahide Mori1

1 Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University Sakyo-ku, Kyoto 606-01 2 Chest Disease Research Institute, Kyoto University Sakyo-ku, Kyoto 606-01 3 Kyoto National Hospital Fushimi-ku, Kyoto, Japan

Correspondence: 4To whom correspondence should be addressed

We examined the presence of cell surface aminopeptidase on cultured porcine granuiosa cells by employing the aminopeptidase assay using alanine-p-nitroanilide and histochemical staining using L-leucyl-β-naphthylamide. Porcine granuiosa cells obtained from follicles 4–5 mm in diameter were cultured for 7 days. The aminopeptidase assay showed that the porcine granuiosa cell culture had aminopeptidase activity and that this activity was inhibited in a dose-dependent manner by bestatin which binds to cell surfaces and inhibits cell surface aminopeptidases. Histochemical staining also indicated that cultured granuiosa cells had aminopeptidase activity. Porcine granuiosa cells were cultured in the presence or absence of porcine follicle stimulating hormone (FSH, 3.125 nmol/1) and/or bestatin (0.4, 4.0 and 40.0 µ/ml) for 7 days, and the production of progesterone and oestradiol was measured. In the presence of porcine FSH, the production of progesterone and oestradiol by granuiosa cells was increased significantly by ~5- and 2-fold respectively. These increases were enhanced further by bestatin (40.0 (µg/ml). In the absence of porcine FSH, progesterone production was enhanced by bestatin (40.0 µg/ml), whereas no significant effect of bestatin on oestradiol secretion was observed. These findings indicate that the inhibition of membrane-bound amino-peptidase(s) on the cell surfaces affects the steroidogenesis of granuiosa cells, and that these aminopeptidase(s) are important regulators of granuiosa cell differentiation.

Key words: bestatin/FSH/granulosa cell/peptidase/steroido-genesis

Submitted on September 25, 1995; accepted on November 30, 1995.


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