Molecular cloning and taste bud-specific expression of a novel cyclic nucleotide-gated channel.

Abstract

Cyclic nucleotide-gated (CNG) channels serve as downstream targets of signaling pathways in vertebrate photoreceptor cells and olfactory sensory neurons. For taste signaling as well, a great deal of information is available predicting the presence of a CNG channel, but no report has been presented on its molecular entity. Here we report on molecular cloning and functional expression of a taste bud-specific CNG channel tentatively named CNGgust. Reverse transcriptase polymerase chain reaction (RT-PCR) primers were synthesized according to some amino acid sequences generally conserved in many CNG channels. RT-PCR was conducted using rat circumvallate papillary mRNA-derived cDNA as a template to obtain positive clones. A corresponding genomic DNA clone was then obtained by screening from a genomic DNA library. Dissecting the entire structure of this gene, we found that the encoding protein had an amino acid sequence similarity of 80% to each of retina and olfactory CNG channels. It was also found by immunostaining with a specific antibody that this gustatory CNG channel (CNGgust) is localized in the tongue and also expressed specifically on the pore side of each taste bud in the circumvallate papillae. Electrophysiological experiments demonstrated that CNGgust resided in a functional state. All these data suggest that CNGgust may be involved in taste signal transduction in sensory cells.