Glutathione S-transferase alpha expressed in porcine Sertoli cells is under the control of follicle-stimulating hormone and testosterone.
Glutathione S-transferases (GSTs) are a family of detoxification isoenzymes present in different tissues including the testis and that conjugate many toxic substrates to glutathione. Among these substrates are carcinogens, mutagens and products of oxidative processes. In the present report we show that GSTalpha is expressed in somatic testicular Leydig cells and Sertoli cells. GSTalpha expression in Sertoli cells is under the hormonal control of FSH, testosterone, and estradiol. In Leydig cells, immunoreactive GSTalpha was present at the neonatal, pubertal, and adult periods. In Sertoli cells, GSTalpha was predominant in pubertal and adult testes (but not in neonatal testes), suggesting that its expression is controlled by gonadotropins. The regulatory action and the mechanisms of action of FSH and testosterone on GSTalpha mRNA and protein levels were studied by using a model of primary cultures of porcine testicular Sertoli cells. FSH increased GSTalpha mRNA levels in a dose-dependent manner (ED50 = 18.5 nm/ml) with a maximal effect observed after 48 h of exposure (a 3-fold increase; P < 0.001). In addition, FSH increased GSTalpha protein, which was detected as a doublet of 28 kDa. Treatment with testosterone enhanced GSTalpha mRNA levels in a dose-dependent (ED50 = 1.4 ng/ml) and time-dependent manner with a maximal effect delayed at 8 h of exposure (a 2-fold increase; P < 0.001). Similarly, Sertoli cell treatment with testosterone metabolites, dihydrotestosterone (DHT) and estradiol, led to an increase in GSTalpha mRNA levels. Because stimulatory effects of FSH and androgens were also observed on GSTalpha protein, we therefore had to determine whether the different hormones were affecting GSTalpha gene transcriptional activity, or GSTalpha mRNA stability, or both. FSH and 8-Br-cAMP (but not testosterone) increased the stability of GSTalpha mRNA. The effects of FSH and testosterone on GSTalpha protein were additive, confirming that both hormones act through distinct mechanisms on the expression of the enzyme. Taken together, the present observations indicate that Sertoli cell GSTalpha is targeted by FSH, testosterone, and its metabolites, and they reinforce the concept that Sertoli cells exert a protective role and are under endocrine control to ward against toxic agents in the context of Sertoli-germ cell interactions during spermatogenesis.
Benbrahim-Tallaa
Lamia
L
Unité 407, Institut National de la Santé et de la Recherche Médicale, Communication Cellulaire en Biologie de la Reproduction, Faculté de Médecine Lyon-Sud, 69921 Oullins Cedex, France.
Tabone
Eric
E
Tosser-Klopp
Gwenola
G
Hatey
François
F
Benahmed
Mohamed
M
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000375:Aging; D000818:Animals; D013196:Dihydrotestosterone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004958:Estradiol / Q000494:pharmacology; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D005982:Glutathione Transferase / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics*; D007700:Kinetics; D007985:Leydig Cells / Q000201:enzymology; D008297:Male; D012333:RNA, Messenger / Q000032:analysis; D012708:Sertoli Cells / Q000201:enzymology*; D013552:Swine; D013737:Testis / Q000254:growth & development; D013739:Testosterone / Q000494:pharmacology*
D007527:Isoenzymes; D012333:RNA, Messenger; D013196:Dihydrotestosterone; D015124:8-Bromo Cyclic Adenosine Monophosphate; D013739:Testosterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone; D005982:Glutathione Transferase; C119995:glutathione S-transferase alpha