Secreted metalloproteinases in testicular cell culture.
It is well known that cultured Sertoli cells secrete plasminogen activators (Lacroix et al., Mol Cell Endocrinol 1977; 9:227-236; Hettle et al., Biol Reprod 1986; 34:895-904). We now show that testicular cells in culture also secrete gelatinolytic metalloproteinases. Gelatin zymographic analysis of concentrated culture medium proteins reveals that Sertoli cells secrete gelatinases of 185 kDa, 110 kDa, 83 kDa, 76 kDa, and 72 kDa in addition to plasminogen activators (PAs). Gelatinase 185 kDa is induced by FSH. Media from Sertoli (epithelial)/peritubular (mesenchymal) cell cocultures contain the Sertoli cell gelatinases and one FSH-stimulated gelatinase of 50 kDa, indicating that gelatinase 50 kDa is regulated by both FSH and cell-cell interactions. A 50-kDa fibronectinolytic activity is also present in the coculture medium from cells grown in the presence of FSH. Casein zymography demonstrates a prominent 30-kDa protease only in media from cocultures. Peritubular cells secrete urokinase-type plasminogen activator (u-PA) and exhibit slight degrading activity at 86 kDa and 74 kDa. The gelatinases are most active in the pH range 7.3-8.5 and are completely or partially inhibited by metal ion chelators indicating that they are metalloproteinases. Our data demonstrate that testicular cells in culture secrete several gelatinases in addition to PAs, and that FSH and coculture conditions regulate some of these secreted proteases. We suggest that the highly regulated secretion of these proteases may well be of physiological importance during testicular development and spermatogenesis.
Sang
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Department of Anatomy and Cell Biology, Georgetown University Medical Center, Washington, District of Columbia 20007.
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Byers
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D000818:Animals; D002478:Cells, Cultured; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D012708:Sertoli Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D013379:Substrate Specificity; D013737:Testis / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism
D005640:Follicle Stimulating Hormone; D008666:Metalloendopeptidases