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Am J Physiol Cell Physiol 274: C1653-C1660, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 6, C1653-C1660, June 1998

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

Y. S. Prakash, Mathur S. Kannan, Timothy F. Walseth, and Gary C. Sieck

Departments of Anesthesiology and of Physiology and Biophysics, Mayo Foundation, Rochester 55905; and Departments of Veterinary PathoBiology, Pediatrics, and Pharmacology, University of Minnesota, St. Paul, Minnesota 55108

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 µM) induced a dose-dependent increase in [Ca2+]i, which was blocked by the cADPR receptor antagonist 8-amino-cADPR (20 µM) and by the RyR blockers ruthenium red (10 µM) and ryanodine (10 µM), 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 µM 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.

ryanodine receptor; second messenger; confocal microscopy; sarcoplasmic reticulum; beta -escin; intracellular calcium concentration


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