ACTIVIN IS PRODUCED BY RAT SERTOLI CELLS-INVITRO AND CAN ACT AS AN AUTOCRINE REGULATOR OF SERTOLI-CELL FUNCTION

The production of activin by Sertoli cells isolated from 21-day-old rats was studied using the mesoderm-inducing activity of activin on Xenopus laevis animal cap explants, immunoprecipitation and Western blotting. Furthermore, the effects of recombinant bovine activin-A on rat Sertoli cell aromatase activity and FSH and androgen receptor gene expression were examined. Animal cap explants from Xenopus laevis blastulas elongated after culture in conditioned medium of Sertoli cells cultured with or without ovine FSH or conditioned medium of the mouse Sertoli cell-derived TM4 cell line. Animal cap explants cultured in control medium remained spherical. This elongation was also found in the more than 10-kilodalton fraction of the conditioned medium and after heating for 10 min at 95 C, indicating that heat-stable activin-like bioactivity is present in the culture medium. Immunoprecipitation of [S-35]methionine-labeled proteins and Western blotting of Sertoli cell-conditioned medium with polyclonal antisera against the inhibin beta-subunits indicated the presence of 24- to 25-kilodalton activin-like immunoreactive material. Sertoli cell aromatase activity was dose-dependently stimulated by ovine FSH after 72 h of culture. Recombinant bovine activin-A partly inhibited this stimulation in a dose-dependent way. This inhibition was also found after 24 h of culture. Furthermore, basal and FSH-stimulated androgen receptor mRNA expression in Sertoli cells and binding of the synthetic androgen R1881 to Sertoli cells were decreased after 24 h of culture in the presence of recombinant bovine activin-A. In the same experiments, FSH receptor mRNA expression was not significantly affected. These results indicate that activin can act as an autocrine regulator of Sertoli cell function.