A factor from bovine granulosa cells preventing oocyte maturation

Abstract

A factor preventing spontaneous dissolution of germinal vesicle (GVBD) of mouse oocytes in vitro was isolated from bovine granulosa cells and designated as granulosa cell factor (GCF). Granulosa cells from medium sized (2-5 mm) follicles were suspended in 10 mM Tris-HCl buffer, pH 8.3, containing 1 M urea and 5 mM EDTA. GCF was separated from the extract by gel filtration on Sephadex G-25 column. The molecular weight (Mr) of GCF was estimated to be less than 6,000 daltons. Oocytes treated with GCF and resuspended in control medium resumed GVBD. The partially purified GCF lost GVBD-preventing activity when digested with pronase but retained its activity when treated with DNase, RNase, or glycosidase. GCF was stable to heating at 100 degrees C for 15 min, not absorbed by charcoal, and did not bind to Concanavalin A-Sepharose 4B. The present findings suggest that GCF is a heat-stable polypeptide and its action is reversible.