Granulosa cells from immature rats primed with diethylstilbestrol (DES) showed a fivefold rise in the activity of secreted plasminogen activator (PA) in response to human FSH. The dose giving 50% of the maximum response (ED50) was 10 ng/ml. Glucocorticoid hormones significantly suppressed PA activity in both control cells and cells stimulated with FSH (ED50 = 50 nmol dexamethasone/l; 1 mumol corticosterone/l). Cortexolone (2.5 mumol/l) significantly ameliorated this suppression, indicating that the response to glucocorticoids is receptor-mediated. These data, together with the time delay required for glucocorticoids to take effect (5 h), suggest that glucocorticoids induce the production of a specific PA inhibitor in granulosa cells.

Two molecular variants of plasminogen activator (PA): urokinase (uPA) and tissue-type plasminogen activator (tPA), have been reported to be synthesized in the rat testis. Data obtained in this study using monospecific antibodies raised against uPA and tPA in immunoblotting and bioimmunoassay protocols consistently demonstrate that only tPA (and not uPA) is synthesized by bovine Sertoli cell-enriched cultures, and is induced by bovine FSH. Zymographic analysis of conditioned medium on gels containing plasminogen and casein showed a dominant PA proteolytic band (72 kDa) which co-migrated with human tPA. A proteolytic band (43 kDa), which was also secreted by FSH-stimulated cells, was not present when protection was afforded from auto-proteolysis by aprotinin, and was therefore concluded to be a proteolytic fragment of tPA, and not uPA.

Plasminogen activation has been shown to be inhibited by the cell-specific production of a number of protease inhibitors belonging to the serine protease inhibitor family. In the bovine testis this inhibitor production is induced by glucocorticoids. Monospecific antibodies raised against the three known classes of plasminogen activator inhibitor were used to identify which type of inhibitor was secreted by bovine Sertoli cell-enriched cultures. Immunoblot analysis and [35S]methionine labelling of newly synthesized proteins revealed that a novel protein with an apparent molecular weight of 49 kDa, which shares antigenic determinants with placental and macrophage PAI and fibroblast protease nexin, is secreted in response to dexamethasone stimulation. This protein was shown by immunoadsorption to be a functionally active inhibitor of both tissue-type and urokinase-type plasminogen activators.

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