title: Effects of FSH and testosterone on highly purified rat Sertoli cells: inhibin alpha-subunit mRNA expression and inhibin secretion are enhanced by FSH but not by testosterone.
abstract: The effects of FSH and testosterone on inhibin mRNA expression and inhibin production by highly purified Sertoli cell preparations were examined. Sertoli cells were isolated from testes of 22-day-old rats by sequential trypsin, collagenase and hyaluronidase treatments, with subsequent osmotic shock treatment on day 3 of culture. Contamination by peritubular and germ cells was less than 0.5 and 1-3% respectively. Intracellular and secreted inhibin levels were measured by radioimmunoassay, using Sertoli cells which were incubated for 24 h in the absence or presence of FSH and testosterone from days 4 to 5 of culture. FSH stimulated the cellular inhibin content and the secreted inhibin level by four- and sevenfold respectively, with a half-maximal effective dose of 5-50 ng/ml. Under the present incubation conditions, testosterone (1 mumol/l) had no effect on immunoreactive inhibin levels in either the presence or absence of FSH. Similarly, the expression of inhibin alpha-subunit mRNA was increased following FSH stimulation, whereas testosterone had no effect. The expression of inhibin beta B-subunit mRNAs was not influenced by FSH or testosterone. It is concluded that highly purified Sertoli cell preparations, with a very low number of peritubular or germ cells, are fully responsive to FSH with respect to inhibin mRNA expression and inhibin production.
authors.affiliation: Department of Biochemistry II, Erasmus University Rotterdam, The Netherlands.
authors.forename: A M
authors.initials: AM
authors.lastname: Toebosch
authors.forename: D M
authors.initials: DM
authors.lastname: Robertson
authors.forename: I A
authors.initials: IA
authors.lastname: Klaij
authors.forename: F H
authors.initials: FH
authors.lastname: de Jong
authors.forename: J A
authors.initials: JA
authors.lastname: Grootegoed
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D007265:Inhibins / Q000096:biosynthesis*; D008297:Male; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000187:drug effects* / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D012333:RNA, Messenger; D013739:Testosterone; D007265:Inhibins; D005640:Follicle Stimulating Hormone