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GENETICALLY MODIFIED ANIMALS AND MODEL ORGANISMS

Amiloride-sensitive NaCl taste responses are associated with genetic variation of ENaC -subunit in mice

¹Section of Oral Neuroscience, Graduate School of Dental Science, Kyushu University, Fukuoka, Japan; ²Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan; and ³Monell Chemical Senses Center, Philadelphia, Pennsylvania

Submitted 15 June 2007 ; accepted in final form 26 October 2007

An epithelial Na⁺ channel (ENaC) is expressed in taste cells and may be involved in the salt taste transduction. ENaC activity is blocked by amiloride, which in several mammalian species also inhibits taste responses to NaCl. In mice, lingual application of amiloride inhibits NaCl responses in the chorda tympani (CT) gustatory nerve much stronger in the C57BL/6 (B6) strain than in the 129P3/J (129) strain. We examined whether this strain difference is related to gene sequence variation or mRNA expression of three ENaC subunits (, β, ). Real-time RT-PCR and in situ hybridization detected no significant strain differences in expression of all three ENaC subunits in fungiform papillae. Sequences of the β- and ENaC subunit genes were also similar in the B6 and 129 strains, but ENaC gene had three single nucleotide polymorphisms (SNPs). One of these SNPs resulted in a substitution of arginine in the B6 strain to tryptophan in the 129 strain (R616W) in the ENaC protein. To examine association of this SNP with amiloride sensitivity of CT responses to NaCl, we produced F₂ hybrids between B6 and 129 strains. Amiloride inhibited CT responses to NaCl in F₂ hybrids with B6/129 and B6/B6 ENaC R616W genotypes stronger than in F₂ hybrids with 129/129 genotype. This suggests that the R616W variation in the ENaC subunit affects amiloride sensitivity of the ENaC channel and provides evidence that ENaC is involved in amiloride-sensitive salt taste responses in mice.

salt taste; amiloride sensitivity; epithelial sodium channel; single nucleotide polymorphisms; nerve recording

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