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Volume 271, Number 36,
Issue of September 6, 1996
pp. 22090-22099
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
Characterization and Hormonal Modulation of Anticoagulant Heparan
Sulfate Proteoglycans Synthesized by Rat Ovarian Granulosa Cells
(Received for publication, February 22, 1996, and in revised form, May 20, 1996)
Ghamartaj
Hosseini
,
Jian
Liu
§
and
Ariane I. de
Agostini
From the Sterility and Gynaecologic Endocrinology
Clinic, Department of Gynaecology and Obstetrics, Geneva University
Hospital, CH 1211 Geneva 14, Switzerland and the
§ Department of Biology, Massachusetts Institute of
Technology, Cambridge, Massachusetts 02139
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. Folliclestimulating 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.

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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
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