Am J Physiol Cell Physiol Journal of Neurophysiology
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Am J Physiol Cell Physiol 264: C803-C809, 1993;
0363-6143/93 $5.00
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AJP - Cell Physiology, Vol 264, Issue 4 C803-C809, Copyright © 1993 by American Physiological Society

ARTICLES

Characterization of endogenous sodium channel gene expressed in Chinese hamster ovary cells

P. H. Lalik, D. S. Krafte, W. A. Volberg and R. B. Ciccarelli
Department of Molecular Biology, Sterling Winthrop Pharmaceuticals Research Division, Rensselaer, New York 12144.

Chinese hamster ovary (CHO-K1) cells were observed to display transient inward Na+ currents of average amplitude (-92 +/- 20 pA), which activated at voltages more than -40 mV, and peak inward currents were observed at potentials equal to or more than +10 mV. Inward Na+ currents in these cells were eliminated after treatment with 500 or 50 nM tetrodotoxin (TTX), whereas 5 nM TTX resulted in 64 +/- 10% inhibition of Na+ current. Using DNA primers designed to bind to the rat brain IIA Na+ channel subtype, we amplified specific polymerase chain reaction (PCR) fragments from CHO-K1 poly-(A)+RNA. The cloning and sequencing of two of these fragments confirmed the presence of an endogenously expressed Na+ channel gene in these cells, which we have termed cho 1. Comparison of the DNA sequence of cho 1 PCR fragments with other known Na+ channel genes indicated a high degree of homology with rat brain Na+ channel subtypes. Northern blots using riboprobes generated from the cho 1 PCR fragments revealed the presence of a specific 9-kb mRNA in these cells. The molecular and electrophysiological data suggest that the cho 1 Na+ channel gene from CHO-K1 cells is closely related to brain-type Na+ channels.


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