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Papers In Press, published online ahead of print February 12, 2003
Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL 35294
Corresponding Author: benos{at}physiology.uab.edu
High-grade glioma cells derived from patient biopsies express an amiloride-sensitive sodium conductance that has properties attributed to the human brain sodium channel family (BNaC), also known as acid-sensing ion channels (ASICs). This amiloride-sensitive conductance was not detected in cells obtained from normal brain tissue, or low-grade or benign tumors. Differential gene profiling data showed that ASIC1 and ASIC2 mRNA were present in normal and low-grade tumor cells. While ASIC1 was present in all high-grade glial cells examined, ASIC2 mRNA was detected in less than half. The main purpose of our work was to examine the molecular mechanisms that may underlie the constitutively activated sodium currents present in high-grade glioma cells. Our results show that 1) gain-of-function mutations of ASIC1 were not present in a number of freshly resected and cultured high-grade gliomas; 2) syntaxin 1A inhibited ASIC currents only when ASIC1 and ASIC2 were coexpressed; and 3) the inhibition of ASIC currents by syntaxin 1A had an absolute requirement for either gamma or delta-hENaC. Transfection of cultured cells originally derived from high-grade gliomas (U87-MG and SK-MG1) with ASIC2 abolished basal amiloride-sensitive sodium conductance; this inhibition was reversed by dialysis of the cell interior with Munc-18, a syntaxin binding protein that typically blocks the interaction of syntaxin with other proteins. Thus, syntaxin 1A cannot inhibit Na+ permeability in the absence of adequate plasma membrane ASIC2 expression, accounting for the observed functional expression of amiloride-sensitive currents in high-grade glioma cells.
J. Biol. Chem, 10.1074/jbc.M300991200
Submitted on January 29, 2003
Revised on February 12, 2003
Accepted on February 12, 2003
Acid-sensing ion channels in malignant gliomas
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