Expression and characterization of a canine hippocampal inwardly rectifying K+ current in Xenopus oocytes

J. Cui, Gail Mandel, D. DiFrancsco, R. P. Kline, P. Pennefather, N. B. Datyner, H. C. Haspel, I. S. Cohen

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Abstract

1. An inwardly rectifying potassium current expressed in Xenopus laevis oocytes injected with canine hippocampal poly(A)+ RNA was investigated with the two-microelectrode voltage clamp technique. 2. Xenopus oocytes injected with canine hippocampal poly(A)+ RNA expressed a current activated by hyperpolarization. This current contained an instantaneous and a time-dependent component. Both components were inwardly rectifying and could be blocked by extracellular Cs+ or Ba2+. 3. The expressed current was carried mainly by K+. Its reversal potential measured in different [K+]os could be fitted by the Nernst equation with a slope of -50.7 per tenfold change in [K+]0 Extracellular Cl and Na+ made minimal contributions to the current. 4. The activation of the expressed current depended on both voltage and [K+]0. Activation started near E(K) and the activation curve shifted along the voltage axis in parallel with E(K) when [K+]0 was altered. 5. The activation time constants of the expressed current also depended on both voltage and [K+]0. The voltage dependence of the time constants was bell-shaped and the peak value was at a potential 30-50 mV more negative than E(K). The voltage dependence of the time constants shifted along the voltage axis when E(K) was changed. 6. The poly(A)+ RNA extracted from canine hippocampus was fractionated in a 10-31% linear sucrose gradient. The size of the mRNA required to express the inwardly rectifying current was estimated to be around 4 kb. 7. In conclusion, the expressed current is an inwardly rectifying potassium current. The canine hippocampal mRNA should be an excellent source for expression-cloning of the inward rectifier channel.

Original languageEnglish (US)
Pages (from-to)229-246
Number of pages18
JournalJournal of Physiology
Volume457
Publication statusPublished - 1992
Externally publishedYes

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ASJC Scopus subject areas

  • Physiology

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Cui, J., Mandel, G., DiFrancsco, D., Kline, R. P., Pennefather, P., Datyner, N. B., ... Cohen, I. S. (1992). Expression and characterization of a canine hippocampal inwardly rectifying K+ current in Xenopus oocytes. Journal of Physiology, 457, 229-246.