Whole-cell currents from the cloned canine cardiac Na+/Ca2+ exchanger NCX1 overexpressed in a fibroblast cell CCL39 |
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Authors: | Uehara A Iwamoto T Shigekawa M Imanaga I |
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Institution: | (1) Department of Physiology, School of Medicine, Fukuoka University, Jonan-ku, Fukuoka 814-80, Japan, JP;(2) Department of Molecular Physiology, National Cardiovascular Center Research Institute, Fujishiro-dai 5, Suita, Osaka 565, Japan, JP |
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Abstract: | A conventional patch-clamp technique was used to record the whole-cell current from the cloned canine cardiac Na+/Ca2+ exchanger NCX1 overexpressed in a fibroblast cell. Ca2+ was extracellularly applied to the Na+-loaded cell to activate the outward current by operating the reverse mode of NCX1. No measurable outward current was ever
elicited from the nontransfected cell. Na+/Ca2+ exchange blocker 5 mM Ni2+ or 3 μM KB-R7943 that was applied extracellularly abolished the outward current. With 140 mM external Li+ (replacing Na+), the outward current was transient during the Ca2+ application. In contrast, with 140 mM external Na+, the outward current was maintained without any inactivation during the Ca2+ application. I–V relations predicted from the whole-cell clamp protocols used were obtained both before and during the Ca2+ application. The exchanger whole-cell currents are thus successfully detectable from NCX1 which is overexpressed in this stable
transfectant system.
Received: 28 February 1997 / Accepted: 9 April 1997 |
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Keywords: | patch-clamp techniques whole-cell recordings heart Na+/Ca2+ exchange |
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