Spatial and temporal characterization of cytoskeletal reorganizations in adherent platelets

Abstract

The functional role of platelets is intricately linked to the dynamic organization of two main components of the cytoskeleton, microtubules and actin fibers. Throughout the phases of platelet activation, spreading, and retraction, both of these essential polymers undergo continuous and orchestrated reorganization. Our investigation of the dynamic cytoskeletal changes during these phases highlights a sequential remodeling of the actin cytoskeleton in adherent platelets from the formation of initial actin nodules through the development of stress fibers and a subsequent return to nodular structures. Concurrently, the marginal ring of microtubules, characteristic of resting platelets, undergoes a re-organization induced by marginal band extension and coiling toward the formation of star-like bundles of microtubules. Subsequently, these bundles are dispersed into individual microtubules, which are re-bundled at later stages before ring-like structures are formed again. These findings suggest a compelling tendency for both cytoskeletal components to revert to their original configurations. Notably, the early steps of platelet cytoskeleton reorganizations have previously been shown to be regulated by the signaling cascade triggered during platelet activation, which leads to an increase of cytosolic calcium concentrations. We show here that later steps are potentially regulated by a progressive decrease of intracellular calcium concentrations as platelets approach the end of their functional lifespan.