Does actin polymerization status modulate Ca2+ storage in human neutrophils? Release and coalescence of Ca2+ stores by cytochalasins

Abstract

The aim of this paper was to establish whether actin polymerization modulated cytosolic Ca2+ storage in human neutrophils. Over the concentration ranges which inhibit actin polymerization, cytochalasins A, B, and D liberated Ca2+ from membrane-bound stores within neutrophils. Two Ca2+ storage sites were identified in neutrophils by the accumulation of the Ca2+ binding probe, chlortetracycline: one at the center of the cell and the other at the cell periphery. Confocal imaging demonstrated that cytochalasins released Ca2+ from the neutrophil periphery, but not from the central Ca2+ store. Ca2+ store release was coupled to Ca2+ influx, suggesting that the peripheral site may be a physiological store containing a Ca2+ influx factor. 3,3'-Dihexyloxacarbocyanine iodide staining organelles, which correlate with Ca2+ release sites, coalesced in neutrophils after treatment with cytochalasins. We propose that peripheral Ca2+ storage sites are restricted from coalescence by cortical polymerized actin and that Ca2+ store coalescence and Ca2+ release are coupled events.