Determination of platelet activation by assaying filamentous actin and detecting membrane alterations.
Thrombin stimulation of human gel-filtered platelets in an unstirred system (without aggregation) results in actin polymerisation. Concomitantly with actin polymerisation the activation markers (CD 63--glycoprotein 53, a 53 kD lysosomal protein and CD 62--GMP 140, a 140 kD alpha granule protein) increase on the platelet membrane as well as the specific binding of monoclonal antibodies to thrombospondin (P 10). Consequently, both assays run in parallel and indicate sensitively platelet activation. Our data also indicate that exposure of subcellular structures following granule fusion is a very early event when platelets are challenged by thrombin and involve a reorganisation of cytoskeletal structures. Cytochalasin E inhibits completely the thrombin-induced actin polymerisation and the platelet aggregation but only partially the thrombin-induced exposure of CD 63. The activation markers CD 62 and P 10 were not influenced.
Spangenberg
P
P
Institute of Pathological Biochemistry, Medical Academy of Erfurt, Germany.
Tschöpe
D
D
Esser
J
J
Schwippert
B
B
Kehrel
B
B
Rösen
P
P
Gries
F A
FA
D000199:Actins / Q000378:metabolism*; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002462:Cell Membrane / Q000378:metabolism; D002850:Chromatography, Gel; D003572:Cytochalasins / Q000494:pharmacology; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors; D006801:Humans; D066298:In Vitro Techniques; D015539:Platelet Activation / Q000187:drug effects*; D013347:Subcellular Fractions / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology
D000199:Actins; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D003572:Cytochalasins; D003850:Deoxyribonuclease I; D013917:Thrombin