Organization of the actin cytoskeleton of resting and activated platelets in suspension.
The present study has employed lysine, phalloidin, and a low concentration of osmic acid to protect the actin cytoskeleton of resting and thrombin-activated platelets during detergent extraction and fixation in suspension. Thin sections of resting platelets revealed a fine amorphous layer containing a few short actin filaments mimicking discoid shape and a randomly dispersed network of actin polymers in the cytoplasm. Following thrombin activation, the cytoskeleton consisted of a peripheral layer of submembrane actin filaments following the contour of shape change, a variable number of filaments in peripheral cytoplasm and extending into pseudopods, and a concentric mass of actin filaments with constricted microtubule rings in cell centers. Prior treatment with cytochalasin B (CB) appeared to reduce the number of actin filaments in resting platelets. Thrombin activation of CB-treated cells resulted in separation of actin filaments, which became concentrated in cell centers, from microtubule coils remaining at the cell periphery. The appearance of detergent-extracted cytoskeletons of platelet actin protected during fixation in suspension by lysine and phalloidin was remarkably similar to that of resting or CB-treated platelets before and after thrombin activation when viewed in conventionally prepared thin sections without exposure to detergent during fixation. The advantage of the new procedure is revelation of the actin filament organization in suspended platelets, which is obscured by the usual glutaraldehyde-osmic acid fixation technique.
Escolar
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Krumwiede
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White
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JG
D000199:Actins / Q000032:analysis*; D000470:Alkaloids; D001792:Blood Platelets / Q000187:drug effects / Q000648:ultrastructure*; D003571:Cytochalasin B; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D004121:Dimethyl Sulfoxide; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods; D017239:Paclitaxel; D013917:Thrombin / Q000494:pharmacology
D000199:Actins; D000470:Alkaloids; D003571:Cytochalasin B; D013917:Thrombin; D017239:Paclitaxel; D004121:Dimethyl Sulfoxide