Organization of von Willebrand factor on surface-activated platelets.
The distribution and organization of von Willebrand factor (vWF) multimers on platelets after surface activation have not been fully characterized. In the present study, washed human platelets were allowed to interact with Formvar-coated, electron microscope grids for 20 minutes at 37 degrees C and then fixed. After fixation, cells were washed and then incubated with buffer alone, human plasma, human plasma preincubated with ristocetin (1.2 mg/mL), purified human vWF plus ristocetin, or bovine plasma. Macromolecular complexes were revealed by ultrastructural immunocytochemistry employing a polyclonal antibody against vWF and protein A-gold (PAG) as the electron-dense probe. vWF multimers were not present in discoid platelets but appeared on the central zone of dendritic cells and over larger central areas of fully spread platelets. Exposure to human plasma alone did not affect the distribution of electron-dense probes for vWF in central regions of surface-activated cells. Incubation of spread platelets with ristocetin-activated human plasma or bovine plasma resulted in the appearance of randomly dispersed, mottled areas of increased density covering the surface from edge to edge. Exposure to vWF antibody and PAG resulted in specific labeling of the dense areas in a serpentine, linear array. The gold-probe distribution suggested that the vWF multimers were not superimposed and were distributed in a random, irregular manner from edge to edge with label-free, clear areas between them. The results extend previous observations demonstrating that glycoprotein Ib-IX receptors are not spontaneously cleared from the plasma membranes of surface-activated platelets by showing that the receptor function of glycoprotein Ib-IX complex remains unchanged.
Escolar
G
G
Servicio de Hemoterapia y Hemostasia, Hospital Clinico y Provincial, Barcelona, Spain.
White
J G
JG
D000818:Animals; D001769:Blood; D001792:Blood Platelets / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D002417:Cattle; D002462:Cell Membrane / Q000502:physiology; D006046:Gold; D006801:Humans; D007150:Immunohistochemistry; D046911:Macromolecular Substances; D015539:Platelet Activation; D010980:Platelet Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology; D013205:Staphylococcal Protein A; D014841:von Willebrand Factor / Q000032:analysis* / Q000737:chemistry
D046911:Macromolecular Substances; D010980:Platelet Membrane Glycoproteins; D013205:Staphylococcal Protein A; D014841:von Willebrand Factor; D012310:Ristocetin; D006046:Gold