Effects of recombinant activin A on in vitro culture of mouse preantral follicles

Activins are part of the intragonadal factors that can modulate the actions of gonadotropins and regulate cellular functions during preantral or early antral stages of folliculogenesis in vivo. In a mouse early preantral follicle culture system, activin A production was measured and recombinant bovine activin A (r-ACT A) was added (10 or 50 ng/ml) to recombinant follicle-stimulating hormone (r-FSH)-supplemented (10 or 100 mIU/ml) medium for a 12-day culture period. Specificity of activin A action was ascertained by addition of recombinant human follistatin (r-FS; 20 or 100 ng/ml). Immunoreactive activin A concentrations in mouse follicle-conditioned medium increased by a factor of 20-50, reaching concentrations from 2 to 5 ng/ml at end of culture. In the initial days of culture, additions of r-ACT A to r-FSH-supplemented medium provoked a dramatic volumetric increase and earlier attachment of the follicle. A dose of 100 ng/ml r-FS was able to block the actions of 10 ng/ml but not those caused by 50 ng/ml r-ACT A. In follicle cultures supplemented with 10 mIU/ml r-FSH, additions of r-ACT induced a dose-dependent inhibin (INH) and estradiol (E-2) increase. Basal and human chorionic gonadotropin (HCG)-induced progesterone (P) production were not influenced by r-ACT A or r-FS additions. Addition of r-ACT A decreased (P = 0.017) the intact follicle survival rate and had no influence on final oocyte diameter. In cultures supplemented by 10 mIU/ml r-FSH, additions of r-ACT A did not influence progression and resumption of meiosis I. Use of a higher r-FSH supplementation dose (100 mlU/ml) tended to affect meiosis I adversely (P = 0.052), and r-ACT A addition amplified this effect significantly (P = 0.007). These in vitro experiments demonstrate pronounced effects from r-ACT on r-FSH-mediated follicle survival, growth, and estrogen biosynthesis. (C) 1998 Wiley-Liss, Inc.