Role of cyclic ADP-ribose in the regulation of [Ca2+]i in porcine tracheal smooth muscle.

The purpose of the present study was to determine whether cyclic ADP-ribose (cADPR) acts as a second messenger for Ca2+ release through ryanodine receptor (RyR) channels in tracheal smooth muscle (TSM). Freshly dissociated porcine TSM cells were permeabilized with beta-escin, and real-time confocal microscopy was used to examine changes in intracellular Ca2+ concentration ([Ca2+]i). cADPR (10 nM-10 microM) induced a dose-dependent increase in [Ca2+]i, which was blocked by the cADPR receptor antagonist 8-amino-cADPR (20 microM) and by the RyR blockers ruthenium red (10 microM) and ryanodine (10 microM), but not by the inositol 1,4,5-trisphosphate receptor blocker heparin (0.5 mg/ml). During steady-state [Ca2+]i oscillations induced by acetylcholine (ACh), addition of 100 nM and 1 microM cADPR increased oscillation frequency and decreased peak-to-trough amplitude. ACh-induced [Ca2+]i oscillations were blocked by 8-amino-cADPR; however, 8-amino-cADPR did not block the [Ca2+]i response to a subsequent exposure to caffeine. These results indicate that cADPR acts as a second messenger for Ca2+ release through RyR channels in TSM cells and may be necessary for initiating ACh-induced [Ca2+]i oscillations.