The broad goal of the proposed experiments is to identify key molecules that allow mammals to detect basic tastes and generate electrical responses that are conducted to brain regions. Molecular mechanisms of taste reception have been a subject of intense investigation over the last 30 years, with great strides made in identifying receptors for bitter, sweet and umami. Much less is known about the nature and function of receptors for sour, the taste that allows us to detect acids in spoiled foods or citrus fruits. In this proposal, we will begin to unravel this problem as we test the contribution of the newly discovered otopetrin proton channels in the transduction of acidic and ionic tastes. These experiments build on the progress made in the last grant application, where we used a combination of cellular, molecular and functional approaches to identify the pH sensitive ion channels in Type III taste receptor cells (TRCs) that mediate sour taste. Notably, we described a novel proton-selective ionic current that is likely to be a key component of sour taste transduction. In the last funding period, we successfully identified the gene that encodes the proton channel, through functional screening of genes enriched in Type III TRCs. Among 41 genes tested, we identified one, encoding the transmembrane protein Otop1 that upon expression induced a proton current in both Xenopus oocytes and HEK-293 cells. Interestingly, Otop1 was first identified as a gene mutated in mice with vestibular defects (?tilted? or tlt) but its function in the vestibular system and elsewhere in the body was not understood. Building on these new results, we propose three specific aims to test the role of the Otop channels in taste signaling.
The first aim will examine the functional distribution of Otop1 across the tongue and palate epithelium, allowing us to answer the question of whether Otop1 is the sole ion channel mediating proton influx in the gustatory system. In the second aim, we will measure cellular responses to acids in wildtype and Otop1 KO mice in order to determine the degree to which Otop1 contributes to sensory responses, ex vivo. In the third aim, we will measure responses from gustatory nerves and assess behavioral thresholds for acid detection in wildtype and Otop1 KO mice to determine the extent to which Otop1 mediates responses to sour taste stimuli in vivo. Together our experiments will allow us to determine if Otop1 functions as a sour taste receptor. Our efforts to identify mechanisms of taste transduction may allow the development of taste modifiers that can be used to enhance palatability of food, reducing the need to add sweeteners that contribute to the development of diabetes or salts that contribute to hypertension. Moreover, the proposed experiments will provide basic information regarding the functional properties of this new family of proton channels that will help us understand their contributions to diverse physiological processes, including brown fat metabolism and the development and maintenance of the vestibular system.
The proposed experiments will use a multi-pronged approach to test the hypothesis that the newly identified Otop1 proton channel is the long sought sour receptor. Otop1 is the founding member of a new family of ion channels, whose functions are largely unknown. Taste plays a defining role in dietary choice, and thus identification of taste receptors has the potential to improve human diet and thereby combat diseases such as obesity or hypertension.
| Tu, Yu-Hsiang; Cooper, Alexander J; Teng, Bochuan et al. (2018) An evolutionarily conserved gene family encodes proton-selective ion channels. Science 359:1047-1050 |
| Ye, Wenlei; Tu, Yu-Hsiang; Cooper, Alexander J et al. (2018) Activation Stoichiometry and Pore Architecture of TRPA1 Probed with Channel Concatemers. Sci Rep 8:17104 |
| Ye, Wenlei; Chang, Rui B; Bushman, Jeremy D et al. (2016) The K+ channel KIR2.1 functions in tandem with proton influx to mediate sour taste transduction. Proc Natl Acad Sci U S A 113:E229-38 |
| Bushman, Jeremy D; Ye, Wenlei; Liman, Emily R (2015) A proton current associated with sour taste: distribution and functional properties. FASEB J 29:3014-26 |
| Liman, Emily R; Zhang, Yali V; Montell, Craig (2014) Peripheral coding of taste. Neuron 81:984-1000 |
| Liman, Emily R (2014) TRP Channels: Pain enters through the side door. Nat Chem Biol 10:171-2 |
