sc-45647 and denatonium-benzoate

The sense of taste comprises at least five distinct qualities: sweet, bitter, sour, salty, and umami, the taste of glutamate. For bitter, sweet, and umami compounds, taste signaling is initiated by binding of tastants to G-protein-coupled receptors in specialized epithelial cells located in the taste buds, leading to the activation of signal transduction cascades. Alpha-gustducin, a taste cell-expressed G-protein alpha subunit closely related to the alpha-transducins, is a key mediator of sweet and bitter tastes. Alpha-gustducin knock-out (KO) mice have greatly diminished, but not entirely abolished, responses to many bitter and sweet compounds. We set out to determine whether alpha-gustducin also mediates umami taste and whether rod alpha-transducin (alpha(t-rod)), which is also expressed in taste receptor cells, plays a role in any of the taste responses that remain in alpha-gustducin KO mice. Behavioral tests and taste nerve recordings of single and double KO mice lacking alpha-gustducin and/or alpha(t-rod) confirmed the involvement of alpha-gustducin in bitter (quinine and denatonium) and sweet (sucrose and SC45647) taste and demonstrated the involvement of alpha-gustducin in umami [monosodium glutamate (MSG), monopotassium glutamate (MPG), and inosine monophosphate (IMP)] taste as well. We found that alpha(t-rod) played no role in taste responses to the salty, bitter, and sweet compounds tested or to IMP but was involved in the umami taste of MSG and MPG. Umami detection involving alpha-gustducin and alpha(t-rod) occurs in anteriorly placed taste buds, however taste cells at the back of the tongue respond to umami compounds independently of these two G-protein subunits.

Topics: Animals; Chorda Tympani Nerve; Crosses, Genetic; Food Preferences; Genotype; Glossopharyngeal Nerve; Glutamates; Guanidines; Inosine Monophosphate; Membrane Glycoproteins; Mice; Mice, Knockout; Quaternary Ammonium Compounds; Quinine; Receptors, Cell Surface; Sodium Glutamate; Sucrose; Taste; Taste Buds; Toll-Like Receptors; Transducin

The transduction of responses to bitter and sweet compounds utilizes guanine nucleotide binding proteins (G proteins) and their coupled receptors. Alpha-gustducin, a transducin-like G protein alpha-subunit, and rod alpha-transducin are expressed in taste receptor cells. Alpha-gustducin knockout mice have profoundly diminished behavioral and electrophysiological responses to many bitter and sweet compounds, although these mice retain residual responses to these compounds. Alpha-gustducin and rod alpha-transducin are biochemically indistinguishable in their in vitro interactions with retinal phosphodiesterase, rhodopsin and G protein betagamma-subunits. To determine if alpha-transducin can function in taste receptor cells and to compare the function of alpha-gustducin versus alpha-transducin in taste transduction in vivo, we generated transgenic mice that express alpha-transducin under the control of the alpha-gustducin promoter in the alpha-gustducin null background. Immunohistochemistry showed that the alpha-transducin transgene was expressed in about two-thirds of the alpha-gustducin lineage of taste receptor cells. Two-bottle preference tests showed that transgenic expression of rod alpha-transducin partly rescued responses to denatonium benzoate, sucrose and the artificial sweetener SC45647, but not to quinine sulfate. Gustatory nerve recordings showed a partial rescue by the transgene of the response to sucrose, SC45647 and quinine, but not to denatonium. These results demonstrate that alpha-transducin can function in taste receptor cells and transduce some taste cell responses. Our results also suggest that alpha-transducin and alpha-gustducin may differ, at least in part, in their function in these cells, although this conclusion must be qualified because of the limited fidelity of the transgene expression.

Topics: Analgesics, Non-Narcotic; Animals; Dose-Response Relationship, Drug; Green Fluorescent Proteins; Guanidines; Immunohistochemistry; Luminescent Proteins; Mice; Mice, Knockout; Mice, Transgenic; Microscopy, Fluorescence; Polymerase Chain Reaction; Promoter Regions, Genetic; Quaternary Ammonium Compounds; Quinine; Sucrose; Sweetening Agents; Taste; Taste Buds; Transducin; Transgenes

Several lines of evidence suggest that both sweet and bitter tastes are transduced via receptors coupled to heterotrimeric guanine-nucleotide-binding proteins (G proteins). Gustducin is a taste receptor cell (TRC)-specific G protein that is closely related to the transducins. Gustducin and rod transducin, which is also expressed in TRCs, have been proposed to couple bitter-responsive receptors to TRC-specific phosphodiesterases to regulate intracellular cyclic nucleotides. Here we investigate gustducin's role in taste transduction by generating and characterizing mice deficient in the gustducin alpha-subunit (alpha-gustducin). As predicted, the mutant mice showed reduced behavioural and electrophysiological responses to bitter compounds, whereas they were indistinguishable from wild-type controls in their responses to salty and sour stimuli. Unexpectedly, mutant mice also exhibited reduced behavioural and electrophysiological responses to sweet compounds. Our results suggest that gustducin is a principal mediator of both bitter and sweet signal transduction.

Topics: Animals; Cloning, Molecular; Facial Nerve; Female; Guanidines; Male; Mice; Mice, Inbred C57BL; Mutagenesis; Quaternary Ammonium Compounds; Quinine; Signal Transduction; Sodium Chloride; Sucrose; Sweetening Agents; Taste; Taste Buds; Transducin