Analysis of Sertoli Cell-Secreted Proteins by Two-Dimensional Gel Electrophoresis

Abstract

The [35S]methionine-labeled proteins secreted from cultured Sertoli cells were analyzed by two-dimensional gel electrophoresis and fluorography. Major polypeptides which were resolved by this procedure were designated by number and further analyzed. Many of these major polypeptides appeared as a series of spots which corresponded to charge isomers. Two of these polypeptides (5 and 6) were shown to be acidic, glycosylated and to comprise the subunits of a dimeric protein of molecular weight 70,000. Some of the polypeptides (4a and 5a) were shown to be secreted from testicular peritubular cells which contaminated the Sertoli cell cultures. However, many of the polypeptides (1,2,3,4,5,5b and 6) were specifically secreted from the Sertoli cells. The fluorogram of the secreted polypeptides obtained from cultured Sertoli cells from 20- or 60-day-old rats were similar to each other but differed from the pattern of polypeptides which were secreted from cultures of Sertoli cells from 10-day-old rats. Polypeptide 3 was identified by immunoprecipitation as testicular transferrin and the synthesis of polypeptide 3 was stimulated when the Sertoli cells were cultured in the presence of follicle-stimulating hormone (FSH), insulin, testosterone and retinol.