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channels in mouse and
rabbit aortic smooth muscle cells: regulation by intracellular
Ca2+ and NO
Vascular Biology Unit, Whitaker Cardiovascular Institute, Boston Medical Center, Boston, Massachusetts 02118
Ca2+-dependent
Cl
(ClCa) channels and their
regulation by intracellular Ca2+
concentration
([Ca2+]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
ClCa currents were activated by
caffeine-induced Ca2+ release.
Single ClCa channels were also
activated by 
200 nM Ca2+ in
inside-out membrane patches and remained active for >5 min in 
1
µM Ca2+ but showed rapid rundown
in 2 mM Ca2+. 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 µM) and
8-(4-chlorophenylthio)-guanosine 3',5'-cyclic monophosphate
(50 µM) did not affect ClCa current, but at a higher concentration SNAP (1 mM) induced a sustained [Ca2+]i
rise, accompanied by a dramatic decrease in caffeine-induced Ca2+ release and
ClCa current. These results
indicate that 1) mouse and rabbit
aortic SMC possess 1.8-pS ClCa channels that are activated by
Ca2+ release from the stores,
2) both activation and rundown of
single ClCa channels depend on
[Ca2+]i,
and 3) NO does not affect
ClCa channels directly or via cGMP
but can inhibit their activation indirectly by decreasing
Ca2+ release from the stores.
single chloride channel; rundown; noise analysis; caffeine-induced calcium release; nitric oxide
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