Identification of sour-taste receptor genes

Abstract

Taste cells located in taste buds respond to gustatory stimuli using a complex arrangement of ion channels, receptor molecules and signaling cascades. Previous electrophysiological experiments have shown that sour taste (essentially a taste of protons) is mediated, at least in part, by apically located amiloride-sensitive channels in the rat. Here, the molecular cloning of sour-taste receptor genes is described. Using a combination of homology screening and functional expression approaches, we screened a rat circumvallate papilla cDNA library and identified acid-sensing ion channel-2a (ASIC2a) and ASIC2b, amiloride-sensitive proton-activated cation channels. In situ hybridization and reverse transcription-polymerase chain reaction experiments showed that ASIC2a and ASIC2b transcripts were localized in taste cells. Immunohistochemical and immunoprecipitation studies revealed that both channels were expressed in a subset of taste cells and that some of the cells expressed ASIC2a/ASIC2b heteromeric assemblies. Immunoelectron microscopic analyses demonstrated that some of the ASIC2a-immunopositive cells had the characteristics of type III cells, which make synaptic contacts with gustatory afferent neurons. According to our electrophysiological studies, stimulation by acetic acid generated larger inward currents in ASIC2a- or ASIC2a/ASIC2b-expressing oocytes than those induced by hydrochloric acid at the same proton concentration and these findings are in good agreement with the well-known taste phenomenon that acetic acid is more sour than hydrochloric acid at equal pH. Taken together, the present results strongly suggest that mucosal protons dissociated from sour-taste substances induce taste cell depolarization through the activation of the ASIC2a and ASIC2a/ASIC2b channels, which leads to transmitter release onto gustatory neurons.