Glycoprotein IIb-IIIa on platelet-derived microparticles, and microparticle structures studied by electron microscopy, confocal laser microscopy and crossed radio-immunoelectrophoresis

Abstract

Shedding of microparticles from the platelet surface is usually associated with exposure of platelet procoagulant activity. Platelet-derived microparticles have been detected in blood in various disease states. In vitro, platelet stimulation with a number of different agonists results in formation of microparticles. In the present study, microparticles induced by platelet stimulation by calcium ionophore or by membrane incorporation of the terminal complement complex C5b-9 were studied using electron microscopy, confocal laser microscopy, flow cytometry and radio-immunoelectrophoresis. When studied by electron microscopy, microparticle morphology was found to be dependent upon the induction method. Platelet stimulation with the calcium ionophore resulted in smaller, more homogeneous and electron dense microparticles than those induced by insertion of the terminal complement complex. With flow cytometry and confocal laser immunofluorescence microscopy, microparticle GPIIb-IIIa was demonstrated using a FITC-conjugated antibody to GPIIIa. Surface-bound GPIIb-IIIa was demonstrated on the microparticles by immunoelectron microscopy. Crossed immunoelectrophoresis of detergent-solubilized microparticles visualized a very prominent GPIIb-IIIa immunoprecipitate arc, and binding of [(125)1]fibrinogen to microparticle GPIIb-IIIa was demonstrated by radio-immunoelectrophoresis. This suggests that the activated GPIIb-IIIa complex is preserved intact during the shedding of microparticles from the platelet surface.