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Department of Pediatrics, University of Iowa, Iowa City, Iowa, 52242
Norepinephrine (NE) increases
Cl
efflux from vascular
smooth muscle (VSM) cells. An increase in
Cl conductance produces
membrane depolarization. We hypothesized that if
Cl currents are important
for agonist-induced depolarization, then interfering with cellular
Cl handling should alter
contractility. Isometric contraction of rat aortic rings was studied in
a bicarbonate buffer. Substitution of extracellular
Cl with 130 mM
methanesulfonate (MS; 8 mM
Cl) did not cause
contraction. NE- and serotonin-induced contractions were potentiated in
this low-Cl buffer, whereas
responses to K+, BAY K 8644, or NE
in the absence of Ca2+ were
unaltered. Substitution of
Cl with
I or
Br suppressed responses to
NE. Inhibition of Cl
transport with bumetanide
(105 M) or bicarbonate-free
conditions (10 mM HEPES) inhibited NE- but not KCl-induced contraction.
The Cl-channel blockers
DIDS (103 M),
anthracene-9-carboxylic acid
(103 M), and niflumic acid
(105 M) all inhibited
NE-induced contraction, whereas tamoxifen
(105 M) did not. Finally,
disruption of sarcoplasmic reticular function with cyclopiazonic acid
(107 M) or ryanodine
(105 M) prevented the
increase in the peak response to NE produced by
low-Cl buffer. We conclude
that a Cl current with a
permeability sequence of I > Br > Cl > MS is
critical to agonist-induced contraction of VSM.
chloride channels; vascular smooth muscle; sodium-potassium-chloride cotransport; chloride/bicarbonate exchange; sarcoplasmic reticulum
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