Hormonal regulation of proteins secreted by cultured pig Sertoli cells: characterization by two-dimensional polyacrylamide gel electrophoresis.
Using a primary culture of immature porcine Sertoli cells, we studied the effect of porcine FSH (pFSH), testosterone and retinoic acid on the labelled secreted protein. Cells were cultured in a chemically defined medium for 3 days and, on day 3, they were incubated for different times in another medium containing labelled amino acids either in the presence or absence of pFSH (50 ng/ml or 2 micrograms/ml), or testosterone (10(-6) M), or retinoic acid (10(-7) M), either alone or in several combinations. After 4 and 8 h of incubation, 20 and 30 secreted peptides were detected respectively by a two-dimensional polyacrylamide gel electrophoresis of the radiolabelled secreted proteins. During these 2 periods, the effect of pFSH was negligible. After 25 h, about 84 spots (pI in the range of 5-8) were identified on the autoradiograms. pFSH (2 micrograms/ml) induced an increase of 14 spots, and a decrease of 7, but at 50 ng/ml only 5 spots were increased and one decreased. Retinoic acid alone induced the increase of one peptide, while testosterone alone or in combination with pFSH (50 ng/ml) did not modify the electrophoretic pattern. When pig Sertoli cells were treated with retinoic acid, testosterone and pFSH (50 ng/ml), the effects on secreted proteins were higher than those induced by pFSH (50 ng/ml) alone.
D000818:Animals; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000032:analysis; D003600:Cytosol / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005640:Follicle Stimulating Hormone / Q000494:pharmacology / Q000502:physiology; D008297:Male; D009698:Nucleoproteins / Q000096:biosynthesis*; D010960:Plasminogen Activators / Q000032:analysis; D012708:Sertoli Cells / Q000378:metabolism*; D013552:Swine; D013739:Testosterone / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D002250:Carbon Radioisotopes; D009698:Nucleoproteins; D013739:Testosterone; D014212:Tretinoin; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP; D010960:Plasminogen Activators