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J. Biol. Chem., Vol. 275, Issue 25, 18887-18896, June 23, 2000
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From the Department of Pharmacology, University of South Alabama
College of Medicine, Mobile, Alabama 36688
Calcium agonists induce membrane depolarization
in endothelial cells through an unknown mechanism. Present studies
tested the hypothesis that pulmonary artery endothelial cells express a
cyclic nucleotide-gated (CNG) cation channel activated by
store-operated calcium entry to produce membrane depolarization. In the
whole-cell configuration, voltage-clamped cells revealed a large
non-inactivating, outwardly rectifying cationic current in the absence
of extra- or intracellular Ca2+ that was reduced upon
replenishment of Ca2+. The inward current was non-selective
for K+, Na+, Cs+, and
Rb+ and was not inhibited by high tetraethylammonium
concentrations. cAMP and cGMP stimulated the current and changed the
cation permeability to favor Na+. Moreover, 8-bromo-cAMP
stimulated the current in voltage-clamped cells in the perforated patch
mode. The cationic current was inhibited by the CNG channel blocker
LY83,583, and reverse transcriptase-polymerase chain reaction cloning
identified expression of a CNG channel resembling that seen in
olfactory neurons. Activation of store-operated calcium entry using
thapsigargin increased a current through the CNG channel. Stimulation
of the current paralleled pulmonary artery endothelial cell membrane
depolarization, and both the current and membrane depolarization were
abolished using LY83,583. Taken together, these data demonstrate
activation of store-operated calcium entry stimulates a CNG channel
producing membrane depolarization. Such membrane depolarization may
contribute to slow feedback inhibition of store-operated calcium entry.
Cyclic Nucleotide-gated Channels Mediate Membrane Depolarization
following Activation of Store-operated Calcium Entry in Endothelial
Cells*
*
This work was supported by American Heart Association
Southeastern Research Consortium fellowships (to S. W. and T. M. M.) and National Institutes of Health Grants HL47063, DK55877 (to M. C.),
DK50151 (to M. L.), HL56050, and HL60024 (to T. S.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Pharmacology,
MSB 3130, University of South Alabama College of Medicine, Mobile, AL
36688. Tel.: 334-460-6010; Fax: 334-460-6798; E-mail: tstevens@jaguar1.usouthal.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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