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J. Biol. Chem., Vol. 280, Issue 43, 36150-36157, October 28, 2005

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Identification and Functional Characterization of a Voltage-gated Chloride Channel and Its Novel Splice Variant in Taste Bud Cells^*

Liquan Huang1, Jie Cao, Hong Wang, Lynn A. Vo, and Joseph G. Brand¶

From the Monell Chemical Senses Center, Philadelphia, Pennsylvania 19104-3308 and the Veterans Affairs Medical Center and the ^¶Department of Biochemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104

Taste bud cells are epithelial cells with neuronal properties. Voltage-dependent ion channels have been physiologically described in these cells. Here, we report the molecular identification and functional characterization of a voltage-gated chloride channel (ClC-4) and its novel splice variant (ClC-4A) from taste bud cells. ClC-4A skipped an exon near its 5'-end, incurring the loss of 60 amino acids at the N terminus. In situ hybridization and immunohistochemistry localized these two channels' transcripts and proteins to a subset of taste bud cells. Electrophysiological recordings of the heterologously expressed channels in Xenopus oocytes showed that ClC-4 and ClC-4A have opposite sensitivity to pH and unique ion selectivity. The chloride channel blockers niflumic acid and 5-nitro-2-(3-phenylpropylamino)benzoic acid had a slight or no inhibitory effect on the conductance of ClC-4, but both blockers inhibited ClC-4A, suggesting that ClC-4A is a candidate channel for an acid-induced 5-nitro-2-(3-phenylpropylamino)benzoic acid-sensitive current. Furthermore, these two channels may play a role in bitter-, sweet-, and umami-mediated taste transmission by regulating transmitter uptake into synaptic vesicles.

Received for publication, July 15, 2005 , and in revised form, August 26, 2005.

^* This work was supported by National Institutes of Health Grant DC05154 (to L. H.), a grant from the United States Department of Veterans Affairs (to J. G. B.), and National Science Foundation Equipment Grant DBI-0216310. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) DQ186662.

¹ To whom correspondence should be addressed: Monell Chemical Senses Center, 3500 Market St., Philadelphia, PA 19104-3308. Tel.: 215-746-0145; Fax: 215-898-2084; E-mail: lhuang{at}monell.org.

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