Immunoelectron-microscopic studies of human platelet thrombospondin, von Willebrand factor, and fibrinogen redistribution during clot formation.
The relative distributions of the human platelet alpha-granule proteins fibrinogen, thrombospondin, and von Willebrand factor were mapped by immunoelectron microscopy in thin cryosections of activated platelets, platelet aggregates, and clots during the first 24 h of in vitro clot formation. In early activated platelets, the results suggest that the canalicular system constitutes a significant component of the external platelet surface, and may act as a compartment for biochemical reactions occurring during granule release. Further, detection of coagulation proteins by various non-morphological procedures may reflect protein contained within canalicular elements. Later in the release process, von Willebrand factor was detected as a major antigen on the platelet canalicular and plasma membranes; thrombospondin, on the other hand, showed minimal binding to platelets and only limited binding to the extensive fibrin network. Comparison of radioimmunoassays of supernatants of thrombin-stimulated platelets in plasma, clotted whole blood, and Triton X-100 platelet releasates indicated that virtually all of the platelet thrombospondin appears in serum. These data confirm the immunocytochemical results indicating that very little platelet thrombospondin binds to the platelet surface, compared with von Willebrand factor, studied here under the same conditions, which binds extensively to the platelet membrane following release and clot formation.
Watkins
S C
SC
Dana-Farber Cancer Institute, Boston, MA.
Raso
V
V
Slayter
H S
HS
D001777:Blood Coagulation / Q000502:physiology*; D001792:Blood Platelets / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D015153:Blotting, Western; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D016253:Microscopy, Immunoelectron; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D011863:Radioimmunoassay; D013048:Specimen Handling; D019699:Thrombospondins; D014841:von Willebrand Factor / Q000737:chemistry / Q000378:metabolism*
D007136:Immunoglobulins; D010980:Platelet Membrane Glycoproteins; D019699:Thrombospondins; D014841:von Willebrand Factor; D005340:Fibrinogen