Evidence of calmodulin involvement in cell volume recovery following hypo-osmotic stress

An influx of Ca2+ into red blood cells of the bivalve mollusc Noetia ponderosa occurs immediately following a hypo-osmotic stress. The volume recovery response to the stress is dependent upon Ca2+]o and is inhibited by phenothiazines. The action of these drugs is on the amino acid regulation portion of the recovery rather than on the ionic portion. Since the phenothiazines are non-specific in action, we have conducted several experiments to decide the site of phenothiazine action on the volume recovery response. The sulfoxide derivatives of both chlorpromazine and trifluoperazine have no effect on volume regulation at the same dose where the parent compound inhibits. At 50-100 times the concentration of the parent compound, the derivatives block both volume regulation and taurine efflux. The phorbol ester, TPA, an activator of protein kinase C, alters the volume recovery, but does so by affecting K+ rather than amino acid regulation. The only phenothiazine target that we can not rule out is calmodulin, which we also demonstrate to be present in the clam red cells. Thus, the data presented suggest that calmodulin is involved in the amino acid regulatory portions of the volume recovery in response to hypo-osmotic swelling.