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Kir2.1 encodes the inward rectifier potassium channel in rat arterial smooth muscle cells
The molecular nature of the strong inward rectifier K + channel in vascular smooth muscle was explored by using isolated cell RT-PCR, cDNA cloning and expression techniques. RT-PCR of RNA from single smooth muscle cells of rat cerebral (basilar), coronary and mesenteric arteries revealed transcripts...
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Published in: | The Journal of physiology 1999-03, Vol.515 (3), p.639-651 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Online Access: | Get full text |
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Summary: | The molecular nature of the strong inward rectifier K + channel in vascular smooth muscle was explored by using isolated cell RT-PCR, cDNA cloning and expression techniques.
RT-PCR of RNA from single smooth muscle cells of rat cerebral (basilar), coronary and mesenteric arteries revealed transcripts
for K ir 2.1. Transcripts for K ir 2.2 and K ir 2.3 were not found.
Quantitative PCR analysis revealed significant differences in transcript levels of K ir 2.1 between the different vascular preparations ( n = 3; P < 0.05). A two-fold difference was detected between K ir 2.1 mRNA and β-actin mRNA in coronary arteries when compared with relative levels measured in mesenteric and basilar preparations.
K ir 2.1 was cloned from rat mesenteric vascular smooth muscle cells and expressed in Xenopus oocytes. Currents were strongly inwardly rectifying and selective for K + .
The effect of extracellular Ba 2+ , Ca 2+ , Mg 2+ and Cs 2+ ions on cloned K ir 2.1 channels expressed in Xenopus oocytes was examined. Ba 2+ and Cs + block were steeply voltage dependent, whereas block by external Ca 2+ and Mg 2+ exhibited little voltage dependence. The apparent half-block constants and voltage dependences for Ba 2+ , Cs + , Ca 2+ and Mg 2+ were very similar for inward rectifier K + currents from native cells and cloned K ir 2.1 channels expressed in oocytes.
Molecular studies demonstrate that K ir 2.1 is the only member of the K ir 2 channel subfamily present in vascular arterial smooth muscle cells. Expression of cloned K ir 2.1 in Xenopus oocytes resulted in inward rectifier K + currents that strongly resemble those that are observed in native vascular arterial smooth muscle cells. We conclude that
K ir 2.1 encodes for inward rectifier K + channels in arterial smooth muscle. |
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ISSN: | 0022-3751 1469-7793 |
DOI: | 10.1111/j.1469-7793.1999.639ab.x |