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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2005;25:2673.)
© 2005 American Heart Association, Inc.

Thrombosis

The Glycoprotein VI-Phospholipase C2 Signaling Pathway Controls Thrombus Formation Induced by Collagen and Tissue Factor In Vitro and In Vivo

Imke C.A. Munnix; Amrei Strehl; Marijke J.E. Kuijpers; Jocelyn M. Auger; Paola E.J. van der Meijden; Marc A.M. van Zandvoort; Mirjam G.A. oude Egbrink; Bernhard Nieswandt; Johan W.M. Heemskerk

From the Departments of Biochemistry (I.C.A.M., M.J.E.K., P.E.J.v.d.M., J.W.M.H.), Biophysics (M.A.M.v.Z.), and Physiology (M.G.A.o.E.), CARIM, Maastricht University, Maastricht, the Netherlands; Rudolf Virchow Center (A.S., B.N.), DFG Research Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany; and Centre for Cardiovascular Sciences Institute of Biomedical Research (J.M.A.), University of Birmingham, Birmingham, United Kingdom.

Correspondence to Johan W.M. Heemskerk, Department of Biochemistry, Maastricht University, PO Box 616, 6200 MD Maastricht, the Netherlands (E-mail jwm.heemskerk{at}bioch.unimaas.nl); or Dr B. Nieswandt, Vascular Biology, Rudolf-Virchow-Zentrum für Experimentelle Biomedizin, Versbacher Str. 9, 97078 Würzburg, Germany (E-mail bernhard.nieswandt@virchow.uni-wuerzburg.de)

Objective— Both collagen and tissue factor can be initiating factors in thrombus formation. We investigated the signaling pathway of collagen-induced platelet activation in interaction with tissue factor–triggered coagulation during the thrombus-forming process.

Methods and Results— In murine blood flowing over collagen, platelet exposure of phosphatidylserine and procoagulant activity, but not adhesion, completely relied on each of the following signaling modules: glycoprotein VI (GPVI), FcR -chain, Src kinases, adaptor protein LAT, and phospholipase C2 (PLC2). On flow in the presence of tissue factor, these signaling components were essential for platelet aggregation and greatly enhanced fibrin clot formation. Collagen-stimulated thrombin generation relied on the presence and activity of GPVI, FcR -chain, Src kinase, LAT, and PLC2. The physiological importance of this GPVI pathway was shown in a FeCl₃-induced in vivo murine thrombosis model. In both venules and arterioles, signaling through GPVI, FcR -chain, and Src kinases enhanced the formation of phosphatidylserine-exposing and fibrin-rich thrombi.

Conclusions— The GPVI-PLC2 activation pathway regulates collagen-dependent coagulation in venous and arterial thrombus formation.

We investigated the contribution of the signaling modules glycoprotein VI, FcR -chain, Src-kinases, LAT, and phospholipase C2 in collagen- and tissue factor–induced thrombus formation. This signaling pathway regulated the formation of procoagulant platelets on collagen, collagen-enhanced thrombin generation, and fibrin clot formation under flow in vitro and in vivo.

Key Words: glycoprotein VI • LAT • platelets • Src kinase • thrombin

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