Regulation of Sertoli cell inhibin production and of inhibin alpha-subunit mRNA levels by specific germ cell types.
To elucidate the endocrine and paracrine regulation of testicular inhibin production, the effects of follicle-stimulating hormone (FSH), (Bu)2cAMP, germ cells (either crude or enriched preparations) and germ cell-conditioned media on inhibin production (immuno- and bio-activities) and the levels of alpha- and beta B-subunit mRNAs were assessed in cultured Sertoli cells isolated from 20-day-old rats. FSH and (Bu)2-cAMP stimulated both secreted and intracellular inhibin levels in a dose-dependent manner. Using cDNA probes corresponding to the alpha-subunit and the beta B-subunit of rat inhibin it was also shown that both FSH and (Bu)2cAMP markedly increased the level of alpha-subunit mRNA but had no effect on the beta B-subunit mRNA. Addition of a crude mixture of germ cells to Sertoli cell monolayers was found to enhance inhibin secretion. Of the different germ cell fractions tested in co-culture, early spermatids reproducibly stimulated both basal and (Bu)2cAMP-induced production of inhibin whereas pachytene spermatocytes only increased the latter; cytoplasts from elongated spermatids (CES) had no effect. Co-culture of Sertoli cells with liver epithelial cells (LEC) significantly enhanced (Bu)2cAMP-induced inhibin levels. Media conditioned by early spermatids consistently and dramatically stimulated the secretion of both bioactive and immunoactive inhibin by Sertoli cells while spent media from pachytene spermatocytes displayed less activity. CES-conditioned media had only minor stimulatory effects, which may have resulted from the contamination of this fraction by spermatids. Media conditioned by LEC had no effect on inhibin production, confirming that the activity of this cell line is not mediated via a diffusible factor. Early spermatids were found to increase levels of the alpha-subunit mRNA. The current study provides evidence for the involvement of germ cells, in particular of early spermatids, in the local testicular regulation of inhibin gene expression and production in the rat. This may be of crucial importance for the ontogeny of this parameter of Sertoli cell function, and has important implications with regard to the postulated endocrine and paracrine roles of inhibin.
Pineau
C
C
Groupe d'Etude de la Fonction de Reproduction chez le Mâle, UA CNRS 256, Université de Rennes I, Campus de Beaulieu, France.
Sharpe
R M
RM
Saunders
P T
PT
Gérard
N
N
Jégou
B
B
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D003470:Culture Media; D015342:DNA Probes; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D007265:Inhibins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism*; D013087:Spermatids / Q000378:metabolism; D013090:Spermatocytes / Q000378:metabolism
D003470:Culture Media; D015342:DNA Probes; D012333:RNA, Messenger; D007265:Inhibins; D003994:Bucladesine; D005640:Follicle Stimulating Hormone