Ca(2+)-dependent Cl(-) Channels in Mouse and Rabbit Aortic Smooth Muscle Cells: Regulation by Intracellular Ca(2+) and NO

Ca(2+)-dependent Cl(-) (Cl(-)(Ca)) channels and their regulation by intracellular Ca(2+) concentration ([Ca(2+)](i)) and nitric oxide (NO) were characterized in mouse and rabbit aortic smooth muscle cells (SMC) using patch clamp and fura 2 imaging. Single channels (1. 8 pS) and whole cell Cl(-)(Ca) currents were activated by caffeine-induced Ca(2+) release. Single Cl(-)(Ca) channels were also activated by >/=200 nM Ca(2+) in inside-out membrane patches and remained active for >5 min in </=1 microM Ca(2+) but showed rapid rundown in 2 mM Ca(2+). Authentic NO or S-nitroso-N-acetylpenicillamine (SNAP) did not affect their activation or rundown in inside-out patches. In the whole cell, SNAP (100 microM) and 8-(4-chlorophenylthio)-guanosine 3',5'-cyclic monophosphate (50 microM) did not affect Cl(-)(Ca) current, but at a higher concentration SNAP (1 mM) induced a sustained [Ca(2+)](i) rise, accompanied by a dramatic decrease in caffeine-induced Ca(2+) release and Cl(-)(Ca) current. These results indicate that 1) mouse and rabbit aortic SMC possess 1.8-pS Cl(-)(Ca) channels that are activated by Ca(2+) release from the stores, 2) both activation and rundown of single Cl(-)(Ca) channels depend on [Ca(2+)](i), and 3) NO does not affect Cl(-)(Ca) channels directly or via cGMP but can inhibit their activation indirectly by decreasing Ca(2+) release from the stores.