Involvement of protein kinase c in the regulation of na/ca exchanger in bovine adrenal chromaffin cells

The Na/Ca exchanger (NCX) exchanges Na+ and Ca bidirectionally through the forward mode (Ca extrusion) or the reverse mode (Ca influx). The present study was undertaken to clarify the role of protein kinase C (PKC) in the regulation of NCX in bovine adrenal chromaffin cells. The Na-loaded cells were prepared by treatment with 100 μmol/L ouabain and 50 μmol/L veratridine. Incubation of Na-loaded cells with Na-free solution in the presence of the Ca channel blockers nicardipine (3 μmol/L) and -conotoxin MVIIC (0.3 μmol/L) caused Ca uptake and catecholamine release. The Na-dependent Ca uptake and catecholamine release were inhibited by 2-[4-[(2,5-difluorophenyl)methoxy] phenoxy]-5-ethoxyaniline (SEA0400; 1 μmol/L) and 2-[2-[4-(4-nitrobenzyloxy) phenyl]isothiourea (KB-R7943; 10 μmol/L), both NCX inhibitors. These results indicate that the Na-dependent responses are mostly due to activation of the NCX working in the reverse mode. In addition, we examined the effects of PKC inhibitors and an activator on the NCX-mediated Ca uptake and catecholamine release. Bisindolylmaleimide I (0.3-10 μmol/L) and chelerythrine (3-100 μmol/L), both PKC inhibitors, inhibited NCX-mediated responses. In contrast, phorbol 12,13-dibutyrate (0.1-10 μmol/L), a PKC activator, enhanced the responses. Bisindolylmaleimide I and chelerythrine, at effective concentrations for inhibition of Na-dependent catecholamine release, had a little or no effect on high K-induced catecholamine release in intact cells or on Ca-induced catecholamine release in β-escin-permeabilized cells. These results suggest that PKC is involved in the activation of NCX in bovine adrenal chromaffin cells. © 2009 Blackwell Publishing Asia Pty Ltd.