CA2-DEPENDENT CL- CURRENT IN CANINE TRACHEAL SMOOTH-MUSCLE CELLS()

Our goal was to investigate the role of Ca2+ entry in regulating Cl- c urrent (I-Cl) in smooth muscle cells from canine trachealis. When stud ies were done using the perforated patch configuration, depolarization elicited a dihydropyridine-sensitive Ca2+ current (I-Ca), followed in many cells by a sustained current. This sustained current reversed di rection close to the Cl- equilibrium potential, consistent with its re presenting I-Cl. The I-Cl was also apparent as slowly deactivating tai l currents seen upon repolarization to negative potentials. The Cl- ch annel blocker niflumic acid abolished both the sustained and tail curr ents, without affecting I-Ca. Several observations indicated that the I-Cl was dependent on Ca2+ entry. I-Cl was increased in magnitude when Ca2+ influx was augmented [by prolonging the depolarization or using BAY K 8644 or acetylcholine (ACh)] and decreased in magnitude when Ca2 + influx was reduced (using nifedipine). Based on these findings, we c onclude that depolarization causes Ca2+ entry, with resultant elevatio n of cytosolic free Ca2+ concentration leading to activation of I-Cl ( I-Cl(Ca)). We investigated whether Ca2+-induced Ca2+ release from the sarcoplasmic reticulum was involved in activation of I-Cl(Ca), by depl eting intracellular stores of Ca2+ using cyclopiazonic acid to block t he sarcoplasmic Ca2+-adenosinetriphosphatase and repeated stimulation with ACh. In such Ca2+-depleted cells, depolarization-mediated Ca2+ en try continued to activate I-Cl(Ca), suggesting that Ca2+ induced Ca2release was not required for its activation. We conclude that Ca2+ ent ry can activate Cl- channels in tracheal smooth muscle. This represent s a positive-feedback system, which would promote excitation and contr action of airway muscle.