Morphological evidence for the association of plasma membrane glycoprotein IIb/IIIa with the membrane skeleton in human platelets.
We examined the association between glycoprotein (GP) IIb/IIIa, a receptor for fibrinogen, and membrane skeletons in both unstimulated and thrombin-activated human platelets. After a treatment with dithiobis succinimidyl propionate (DTSP), a cross-linker, unstimulated and activated platelets were simultaneously extracted and fixed with a fixing solution containing Triton X-100. Also, the localization of GPIIb/IIIa on the plasma membrane was observed by a preembedding staining method of unextracted platelets. In unstimulated platelets, 20-40% of the whole plasma membrane remained in the detergent-extracted samples. Amorphous structures with 10-70 nm in diameters are distributed at 20 to 100-nm intervals on the surface of plasma membrane. Similar structures also were identified in the intact platelets by the immunocytochemical method. By careful inspection, we found that most of the amorphous structures that contained gold particles were connected to the submembrane zone just beneath the plasma membrane. The submembrane zone was identified as the membrane skeleton because actin was detected in the zone. After activation, detergent-insoluble granules were surrounded by dense networks of microfilaments in the central part of platelets. The filaments were identified as actin and became associated with myosin. These results demonstrate that GPIIb/IIIa on the plasma membrane is connected to the membrane skeleton and suggest that, during activation, actin filaments which extend into the cytoplasm from the membrane skeleton increase and form dense networks around Triton-insoluble granules.
Department of Cardiovascular Research, Tokyo Metropolitan Institute of Medical Science, Japan.
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H
Suzuki
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K
Tanoue
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H
Yamazaki
D000199:Actins / Q000032:analysis; D001792:Blood Platelets / Q000648:ultrastructure*; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure; D003599:Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D009218:Myosins / Q000032:analysis; D015539:Platelet Activation / Q000502:physiology; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*
D000199:Actins; D010980:Platelet Membrane Glycoproteins; D009218:Myosins