Characterization and hormonal modulation of anticoagulant heparan sulfate proteoglycans synthesized by rat ovarian granulosa cells

Abstract

Anticoagulant heparan sulfate proteoglycans endow the vascular endothelium with antithrombotic properties, but their role outside the vascular bed is unknown. Granulosa cells form an avascular compartment in the ovarian follicle, in which a heparin-like activity has been described. At ovulation extravascular coagulation occurs around ovulatory follicles, and after expulsion of the oocyte, a fibrin clot forms in the antral cavity. Granulosa cells synthesize two major heparan sulfate proteoglycans, whose anticoagulant nature has not been investigated. The purpose of this study was to characterize anticoagulant heparan sulfate proteoglycans synthesized by rat ovarian granulosa cells. Affinity purified 35S-labeled anticoagulant heparan sulfate glycosaminoglycans represent 6.5% of the total heparan sulfate synthesized, and they contain 13% 3-O-sulfated disaccharides that are markers of the antithrombin-binding site of heparin. The biological activity of granulosa cell heparan sulfate proteoglycans was demonstrated by their ability to bind antithrombin and to accelerate the formation of thrombin-antithrombin complexes. The impact of hormonal stimulation on granulosa cell anticoagulant heparan sulfate proteoglycans was studied using 125I-antithrombin binding assays. Follicle-stimulating hormone induced a redistribution of anticoagulant heparan sulfate proteoglycans from the granulosa cell layer to the culture medium, indicating that their distribution could be modulated according to the stage of follicular development. These results suggest that anticoagulant heparan sulfate might be critically located in the follicle to maintain fluidity around the oocyte until its expulsion at ovulation.