Three-dimensional organization of the platelet cytoskeleton during adhesion in vitro: observations on human and nonhuman primate cells

Abstract

Adhesion of platelets in vitro resulted in rapid polymerization of the amorphous cytoplasmic ground substance into an organized cytoskeletal superstructure. This cytoskeleton, characterized through the use of whole-mount and stereo (3-D), high-voltage microscopy in conjunction with morphometrics and cytochemistry, comprised four major size classes of filaments organized in distinctive zones. The central matrix, or granulomere, at the center of the cell mass, was an ill-defined meshwork of 80-100-A filaments which enshrouded granules, dense bodies, and elements of the dense tubular system as identified through peroxidase cytochemistry. Demarcating this central matrix was a trabecular zone containing 30-50, 80-100, and 150-170 A filaments in an open and rigid-appearing lattice. Circumscribing the trabecular zone and extending to the margins of the hyalomere was the third region, the peripheral web, in which 70-A filaments were arranged in a tight honeycomb lattice. This organizational pattern was retained in cytoskeletons prepared by Triton x-100 extraction of the adherent cells, and was observed in basally located cells of aggregates which formed subsequent to adhesion. Our observations are consistent with biochemical studies of cytoskeletons prepared from suspended platelets and suggest a contractile protein composition for the superstructure during adhesion.