Endogenous TRPM4-like channel in Chinese hamster ovary (CHO) cells

Abstract

Chinese hamster ovary (CHO) cells used in many transfection studies have been found to endogenously express channels permeable to monovalent cations, but not to divalent cations. In the presence of intracellular Ca2+, 23-pS channel with a linear current-voltage (I-V) relationship could be frequently observed in inside-out patches but not in cell-attached patches. The open probability was voltage-dependent, which is higher at positive potentials. The channel was dose-dependently activated by relatively high level of Ca2+ (EC50 = 1.04 +/- 0.08 mM), and sensitively inhibited by 100 mu M ATP, ADP, AMP, and I mM spermine. However, ruthenium red (2 mu M) had no effect. Reverse transcript polymerase chain reaction (RT-PCR) supported the presence of mRNA encoding TRPM4b channel protein. Western blot assay finally confirmed the presence of this channel protein in membrane fraction of CHO cells. These results provide evidence that CHO cells express an endogenous TRPM4b-like channel, and thereby can be used as a tool to study de novo regulation/modulation of TRPM4 channel. (c) 2008 Elsevier Inc. All rights reserved.