Platelet cytoskeleton: immunofluorescence studies on thrombin-activated platelets.
Cytoskeletal proteins were isolated from chicken gizzard smooth muscle and from platelets and antibodies prepared against them. It was shown by indirect immunofluorescence technique that actin, alpha-actinin, and vinculin are not present on the surface of platelets. Physiologic concentrations of thrombin (0.04 to 0.5 U/ml) that induce platelet aggregation and release in the presence of calcium from freshly isolated platelets do not induce platelet changes resulting in the availability of the cytoskeleton to antibodies. Because the F(ab')2 fragments of anti-cytoskeletal proteins IgG do not inhibit thrombin-induced aggregation of platelets, the direct role of these proteins in thrombin-induced platelet aggregation, as with ADP and collagen, may be rejected. However, when freshly isolated platelets are treated with thrombin (1 U/ml), antibodies to actin, alpha-actinin, and vinculin stained the platelets; therefore, this demonstrates that thrombin at this high and probably nonphysiologic concentration induces a reorganization of the membrane components with the subsequent exposure of the proteins of the cytoskeleton. We demonstrate interaction between isolated actin and alpha-actinin but not vinculin with fibronectin. After stimulation of platelets by thrombin, certain cytoskeletal proteins may interact with subendothelial fibronectin and thereby promote and consolidate platelet adhesion.
D000818:Animals; D000918:Antibody Specificity; D001792:Blood Platelets / Q000032:analysis* / Q000187:drug effects / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000097:blood*; D003599:Cytoskeleton / Q000648:ultrastructure*; D005353:Fibronectins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits / Q000276:immunology; D013917:Thrombin / Q000502:physiology*
D003598:Cytoskeletal Proteins; D005353:Fibronectins; D007140:Immunoglobulin Fab Fragments; D013917:Thrombin