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

J. Cui, G. Mandel, D. DiFrancesco, 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+]o. Extracellular Cl‐ and Na+ made minimal contributions to the current. 4. The activation of the expressed current depended on both voltage and [K+]o. Activation started near EK and the activation curve shifted along the voltage axis in parallel with EK when [K+]o was altered. 5. The activation time constants of the expressed current also depended on both voltage and [K+]o. The voltage dependence of the time constants was bell‐shaped and the peak value was at a potential 30‐50 mV more negative than EK. The voltage dependence of the time constants shifted along the voltage axis when EK 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
Pages (from-to)229-246
Number of pages18
JournalThe Journal of Physiology
Volume457
Issue number1
DOIs
StatePublished - Nov 1 1992

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