Involvement of IP3-receptor activation in endothelin-1-induced Ca2+ influx in rat pulmonary small artery

We examined the endothelin-1 (ET-1)-inducal increase in the intracellular free Ca2+ concentration ([Ca2+](i)) in fura-2-loadecl rat pulmonary small arteries. ET-1 (30 nM) elicited a long-lasting increase in [Ca2+](i) in physiological salt solution (PSS). In subsequent experiments, arteries were pretreated with BQ-788, an ETB-specific blocker, to allow us to focus on responses mediated via the ETA receptor, the existence of which was confirmed by immunohistochemistry. In Ca2+-free PSS, ET-1 evoked a small transient increase in [Ca2+]i indicating Ca2+ release from the SR (sarcoplasmic reticulum). After a switch to PSS (containing 2 mM CaCl2), ET-1 elicited a long-lasting increase in [Ca2+](i); that was not inhibited by 1 mu M nicardipine, an L-type Ca2+-channel inhibitor, suggesting involvement of a Ca2+-influx pathway independent of that channel. In arteries preincubated with 30 mu M cyclopiazonic acid (CPA) or 2 mu M thapsigargin (TG), the ET-1-induced Ca2+-release was greatly reduced, and the induced Ca2+-influx was attenuated. U-73122, a phospholipase C (PLC) inhibitor, had inhibitory effects similar to those of CPA and TG on the ET-1-induced Ca2+-release and Ca2+-influx, whereas U-73343, an inactive analogue of U-73122, had no such effects. Two putative membrane permeable IP3 receptor blockers, 2-aminoethoxydiphenyl borate (2APB, 50 mu M) and Xestospongin C(20 mu M), (a) almost completely inhibited the ET-1-induced Ca2+-release and Ca2+-influx, and (b) reduced the ET-1-induced contraction. These results indicate that in rat pulmonary small arteries, ET-1 induces receptor operated Ca2+ influx via the ETA receptor, and that this influx interacts with InsP(3)-receptor activation. (C) 2013 Elsevier B.V. All rights reserved.