title: Transglutaminase activity in testicular homogenates and serum-free Sertoli cell cultures.
abstract: Transglutaminase (EC 2.3.2.13) (TGase) activity has been localized in homogenates of rat Leydig cells and seminiferous tubules and is present in cytosol and membrane fractions. The enzyme has a requirement for Ca2+ and when the acceptor substrate casein was deleted from the assay mixture, incorporation of [14C]putrescine into cytosolic and membrane fractions occurred. Transglutaminase was also detected in Sertoli cells cultured in serum-free medium. Sertoli cells reside within the seminiferous tubule and are involved in normal spermatogenesis. Sertoli cell TGase has a strict requirement for Ca2+ and is not activated by Mg2+. Activation of the enzyme occurs with as little as 0.3 microM Ca2+; however, consistent with intracellular calcium levels, maximum stimulation occurred at 1.9 mM Ca2+. Sertoli cell TGase activity is markedly stimulated if the cells are cultured in 10% fetal bovine serum rather than in serum-free medium. Inhibition of Sertoli cell TGase by monodansylcadaverine concomitantly decreased the response of the cells to follicle-stimulating hormone (FSH)-induced secretion of cAMP but did not change basal cAMP levels. These data suggest that TGase may play a facilitative rather than an absolute role in activation of Sertoli cells by FSH and the resultant secretion of cellular products. This may occur through modulation of activities of membrane and cytosolic components by TGase.
authors.forename: J A
authors.initials: JA
authors.lastname: Dias
mesh_terms: D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D009994:Osmolar Concentration; D051381:Rats; D011919:Rats, Inbred Strains; D012671:Seminiferous Tubules / Q000201:enzymology; D012708:Sertoli Cells / Q000166:cytology / Q000201:enzymology*; D013737:Testis / Q000201:enzymology*; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D003470:Culture Media; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP; D011503:Transglutaminases; D002122:Calcium Chloride