Lysophosphatidic acid opens a Ca(++) channel in human erythrocytes

Lysophosphatidic acid (LPA) is a lipid-derived second messenger that mobilizes many cells of the circulatory and vascular systems to assist in thrombus development and wound healing. LPA, however, has not been tested on human erythrocytes, largely because erythrocytes are considered to be both biologically inert and inactive in intercellular communication. To test this presumption, we have examined the impact of LPA on signaling reactions within the human red blood cell (RBC). Using both (45)Ca(++) and a Ca(++)-sensitive fluorescent probe (Fluo-3), we demonstrated that LPA, but not phosphatidic acid or the closely related sphingosine-1-phosphate, stimulates the influx of micromolar quantities of extracellular Ca(++) into fresh RBCs. This Ca(++) influx was shown to be channel mediated rather than leak promoted because the influx was observed at LPA concentrations too low to perturb membrane integrity, it was inhibited by P-type but not L-type Ca(++) channel blockers, it was inhibited by broad-specificity protein kinase inhibitors, and it was not induced by inactive analogues of LPA. Further characterization reveals that only approximately 25% of the RBCs participate in LPA-induced Ca(++) entry and that within this active population, Ca(++) gating occurs in an all-or-nothing manner. Because the stimulation of Ca(++) uptake occurs at LPA concentrations (1-5 micromol/L) known to occur near a developing thrombus and because the internalized Ca(++) can potentially promote prothrombic properties in the stimulated RBCs, we conclude that RBCs are not insensitive to signals released from other cells.