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Stimulating Effect of Both Human Recombinant Inhibin A and Activin A on Immature Porcine Leydig Cell Functions in Vitro¹

INSERM-INRA U-418 and IFR D’Endocrinologie, Hôpital Debrousse (H.L., F.C., P.S., P.D., J.M.S.), 69322 Lyon; and Clinique Endocrinologique, Hospices Civils de Lyon, Hôpital de l’Antiquaille (H.L.), 69321 Lyon, France; and Genentech (J.P.M.), South San Francisco, California 94080

Address all correspondence and requests for reprints to: Dr. J. M. Saez, INSERM-INRA U-418, Hôpital Debrousse, 69322 Lyon, France. E-mail: pdurand{at}univ-lyon1.fr

In addition to the regulation of FSH secretion, it has been clearly shown that inhibin and activin have paracrine/autocrine effects in the gonads. We have studied the effect of human recombinant inhibin A and human recombinant activin A on immature porcine Leydig cells in vitro. Leydig cells were prepared by collagenase digestion of testes from 3-week-old piglets, purified on Percoll gradient, then cultured in a chemically defined medium. The cells were treated with increasing amounts of inhibin A or activin A (0.5–200 ng/ml). Direct application of either inhibin A or activin A on Leydig cells for 4 or 48 h did not stimulate basal testosterone secretion. Conversely, treatment of the cells for 48 h with either factor resulted in a dose-dependent increase in hCG-stimulated testosterone secretion (10^-9 M hCG, 2 h) with a maximal effect of 2.40 ± 0.37- and 2.43 ± 0.37-fold increases for inhibin A and activin A, respectively, and these changes were associated with a slight increase in LH/hCG-binding sites (1.37 ± 0.19- and 1.24 ± 0.11-fold increases). In addition, both inhibin A and activin A enhanced messenger RNA (mRNA) levels of LH/hCG receptor (2.75 ± 0.40- and 2.53 ± 0.60-fold increases) and cytochrome P450 17-hydroxylase (6 ± 1- and 3.5 ± 0.6-fold increases), but had no effect on side-chain cleavage cytochrome P450 or cytochrome P450 aromatase mRNAs. 3ß-Hydroxysteroid dehydrogenase mRNA levels were increased (3.1 ± 1.3-fold increase) by activin A, but not by inhibin A. However, inhibin A blocked the stimulatory action of activin A. In keeping with these changes in the steroidogenic enzyme mRNAs, both peptides enhanced the conversion of exogenous 22R-hydroxycholesterol and progesterone, but only activin A increased the conversion of dehydroepiandrosterone into testosterone. In conclusion, our findings demonstrate that both inhibin A and activin A have a stimulatory effect on immature porcine Leydig cell differentiated function in vitro. As inhibin has a stimulatory and activin has an inhibitory effect on rat Leydig cell function in vitro, the effects of these factors on Leydig cells seem to be species dependent.

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