Abstract
We used differential screening of cDNAs from individual taste receptor cells to identify candidate taste transduction elements in mice. Among the differentially expressed clones, one encoded Trpm5, a member of the mammalian family of transient receptor potential (TRP) channels. We found Trpm5 to be expressed in a restricted manner, with particularly high levels in taste tissue. In taste cells, Trpm5 was coexpressed with taste-signaling molecules such as α-gustducin, Gγ13, phospholipase C-β2 (PLC-β2) and inositol 1,4,5-trisphosphate receptor type III (IP3R3). Our heterologous expression studies of Trpm5 indicate that it functions as a cationic channel that is gated when internal calcium stores are depleted. Trpm5 may be responsible for capacitative calcium entry in taste receptor cells that respond to bitter and/or sweet compounds.
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References
Gilbertson, T.A., Damak, S. & Margolskee, R.F. The molecular physiology of taste transduction. Curr. Opin. Neurobiol. 10, 519–527 (2000).
Lindemann, B. Receptors and transduction in taste. Nature 413, 219–225 (2001).
Adler, E. et al. A novel family of mammalian taste receptors. Cell 100, 693–702 (2000).
Matsunami, H., Montmayeur, J.P. & Buck, L.B. A family of candidate taste receptors in human and mouse. Nature 404, 601–604 (2000).
Chandrashekar, J. et al. T2Rs function as bitter taste receptors. Cell 100, 703–711 (2000).
Bachmanov, A.A. et al. Positional cloning of the mouse saccharin preference (Sac) locus. Chem. Senses 26, 925–933 (2001).
Nelson, G. et al. Mammalian sweet taste receptors. Cell 106, 381–390 (2001).
Sainz, E., Korley, J.N., Battey, J.F. & Sullivan, S.L. Identification of a novel member of the T1R family of putative taste receptors. J. Neurochem. 77, 896–903 (2001).
Max, M. et al. Tas1r3, encoding a new candidate taste receptor, is allelic to the sweet responsiveness locus. Sac. Nat. Genet. 28, 58–63 (2001).
Montmayeur, J.P., Liberles, S.D., Matsunami, H. & Buck, L.B. A candidate taste receptor gene near a sweet taste locus. Nat. Neurosci. 4, 492–498 (2001).
Kitagawa, M., Kusakabe, Y., Miura, H., Ninomiya, Y. & Hino, A. Molecular genetic identification of a candidate receptor gene for sweet taste. Biochem. Biophys. Res. Commun. 283, 236–242 (2001).
Chaudhari, N., Landin, A.M. & Roper, S.D. A metabotropic glutamate receptor variant functions as a taste receptor. Nat. Neurosci. 3, 113–119 (2000).
Nelson, G. et al. An amino-acid taste receptor. Nature 416, 199–202 (2002).
McLaughlin, S.K., McKinnon, P.J. & Margolskee, R.F. Gustducin is a taste-cell–specific G protein closely related to the transducins. Nature 357, 563–569 (1992).
Wong, G.T., Gannon, K.S. & Margolskee, R.F. Transduction of bitter and sweet taste by gustducin. Nature 381, 796–800 (1996).
Huang, L. et al. Gγ13 colocalizes with gustducin in taste receptor cells and mediates IP3 responses to bitter denatonium. Nat. Neurosci. 2, 1055–1062 (1999).
Rossler, P. et al. G protein βγ complexes in circumvallate taste cells involved in bitter transduction. Chem. Senses. 25, 413–421 (2000).
Rossler, P., Kroner, C., Freitag, J., Noe, J. & Breer, H. Identification of a phospholipase C β subtype in rat taste cells. Eur. J. Cell. Biol. 77, 253–261 (1998).
Clapp, T.R., Stone, L.M., Margolskee, R.F. & Kinnamon, S.C. Immunocytochemical evidence for coexpression of Type III IP3 receptor with signaling components of bitter taste transduction. B. M. C. Neurosci. 2, 6 (2001).
Miyoshi, M.A., Abe, K. & Emori, Y. IP3 receptor type 3 and PLCβ2 are coexpressed with taste receptors T1R and T2R in rat taste bud cells. Chem. Senses 26, 259–265 (2001).
Misaka, T. et al. Taste buds have a cyclic nucleotide-activated channel, CNGgust. J. Biol. Chem. 272, 22623–22629 (1997).
Bernhardt, S.J., Naim, M., Zehavi, U. & Lindemann, B. Changes in IP3 and cytosolic Ca2+ in response to sugars and non-sugar sweeteners in transduction of sweet taste in the rat. J. Physiol. 490, 325–336 (1996).
Ogura, T., Mackay-Sim, A. & Kinnamon, S.C. Bitter taste transduction of denatonium in the mudpuppy Necturus maculosus. J. Neurosci. 17, 3580–3587 (1997).
Ming, D., Ruiz-Avila, L. & Margolskee, R.F. Characterization and solubilization of bitter-responsive receptors that couple to gustducin. Proc. Natl. Acad. Sci. USA 95, 8933–8938 (1998).
Putney, J.W. Jr. & McKay, R.R. Capacitative calcium entry channels. Bioessays 21, 38–46 (1999).
Clapham, D.E., Runnels, L.W. & Strubing, C. The trp ion channel family. Nat. Rev. Neurosci. 2, 387–396 (2001).
Wong, G.T., Ruiz-Avila, L. & Margolskee, R.F. Directing gene expression to gustducin-positive taste receptor cells. J. Neurosci. 19, 5802–5809 (1999).
Yatsuki, H. et al. Sequence-based structural features between Kvlqt1 and Tapa1 on mouse chromosome 7F4/F5 corresponding to the Beckwith-Wiedemann syndrome region on human 11p15.5: long-stretches of unusually well conserved intronic sequences of kvlqt1 between mouse and human. DNA Res. 7, 195–206 (2000).
Enklaar, T. et al. Mtr1, a novel biallelically expressed gene in the center of the mouse distal chromosome 7 imprinting cluster, is a member of the Trp gene family. Genomics 67, 179–187 (2000).
Prawitt, D. et al. Identification and characterization of MTR1, a novel gene with homology to melastatin (MLSN1) and the trp gene family located in the BWS-WT2 critical region on chromosome 11p15.5 and showing allele-specific expression. Hum. Mol. Genet. 9, 203–216 (2000).
Montell, C. et al. A unified nomenclature for the superfamily of TRP cation channels. Mol. Cell. 9, 229–231 (2002).
Paulsen, M. et al. Sequence conservation and variability of imprinting in the Beckwith-Wiedemann syndrome gene cluster in human and mouse. Hum. Mol. Genet. 9, 1829–1841 (2000).
Gillo, B. et al. Coexpression of Drosophila TRP and TRP-like proteins in Xenopus oocytes reconstitutes capacitative Ca2+ entry. Proc. Natl. Acad. Sci. USA 93, 14146–14151 (1996).
Lytton, J., Westlin, M. & Hanley, M.R. Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. J. Biol. Chem. 266, 17067–17071 (1991).
Bobanovic, L.K. et al. Molecular cloning and immunolocalization of a novel vertebrate trp homologue from Xenopus. Biochem. J. 340, 593–599 (1999).
Halaszovich, C.R., Zitt, C., Jungling, E. & Luckhoff, A. Inhibition of TRP3 channels by lanthanides. Block from the cytosolic side of the plasma membrane. J. Biol. Chem. 275, 37423–37428 (2000).
Lomax, R.B., Herrero, C.J., Garcia-Palomero, E., Garcia, A.G. & Montiel, C. Capacitative Ca2+ entry into Xenopus oocytes is sensitive to omega-conotoxins GVIA, MVIIA and MVIIC. Cell Calcium 23, 229–239 (1998).
Ogura, T. Acetylcholine increases intracellular Ca2+ in taste cells via activation of muscarinic receptors. J. Neurophysiol. 87, 2643–2649 (2002).
Ogura, T., Margolskee, R.F. & Kinnamon, S.C. Taste receptor cell responses to the bitter stimulus denatonium involve Ca2+ influx via store-operated channels. J. Neurophysiol. 87, 3152–3155 (2002).
Randriamampita, C. & Tsien, R.Y. Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature 364, 809–814 (1993).
Birnbaumer, L. et al. Mechanism of capacitative Ca2+ entry (CCE): interaction between IP3 receptor and TRP links the internal calcium storage compartment to plasma membrane CCE channels. Recent. Prog. Horm. Res. 55, 127–161 (2000).
Hofmann, T. et al. Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol. Nature 397, 259–263 (1999).
Yan, W. et al. Bitter taste transduced by PLC-β(2)-dependent rise in IP3 and alpha-gustducin-dependent fall in cyclic nucleotides. Am. J. Physiol. Cell. Physiol. 280, C742–C751 (2001).
Hofmann, T., Schaefer, M., Schultz, G. & Gudermann, T. Subunit composition of mammalian transient receptor potential channels in living cells. Proc. Natl. Acad. Sci. USA 99, 7461–7466 (2002).
Hardie, R.C. & Minke, B. The trp gene is essential for a light-activated Ca2+ channel in Drosophila photoreceptors. Neuron 8, 643–651 (1992).
Liman, E.R., Corey, D.P. & Dulac, C. TRP2: a candidate transduction channel for mammalian pheromone sensory signaling. Proc. Natl. Acad. Sci. USA 96, 5791–5796 (1999).
Stortkuhl, K.F., Hovemann, B.T. & Carlson, J.R. Olfactory adaptation depends on the Trp Ca2+ channel in Drosophila. J. Neurosci. 19, 4839–4846 (1999).
Liedtke, W. et al. Vanilloid receptor-related osmotically activated channel (VR-OAC), a candidate vertebrate osmoreceptor. Cell 103, 525–535 (2000).
Caterina, M.J. et al. Impaired nociception and pain sensation in mice lacking the capsaicin receptor. Science 288, 306–313 (2000).
Chan, K.W. et al. A recombinant inwardly rectifying potassium channel coupled to GTP-binding proteins. J. Gen. Physiol. 107, 381–397 (1996).
Acknowledgements
We are grateful to M. Cahalan, D. Logothetis and S. Kinnamon for critical reading of the manuscript. R.F.M. is an Associate Investigator of the Howard Hughes Medical Institute. This research was supported by grants from the U.S. National Institutes of Health: DC03055 and DC03155 (R.F.M.), MH57241 (M.M.) and DC00310 (L.H.).
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Supplementary Fig. 1.
Identification of differentially expressed cDNAs in taste receptor cells. Individual clones picked from a cDNA library generated from a single α-gust+ TRC were arrayed in duplicate on nylon membranes then hybridized in parallel with radiolabeled probes from an α-gust+ (Membrane 1) or an α-gust− (Membrane 2) taste cell. The first row of each membrane contained the following cDNAs as internal standards: Gβ1 (A1/A'1), a serial dilution of G3PDH (A3/A'3, A4/A'4, A5/A'5, A6/A'6: 50, 5, 0.5, 0.05 ng, respectively), RGS2 (A7/A'7), α-gustducin (A8/A'8, A9/A'9: 50, 5 ng, respectively), cytokeratin 8 (A10/A'10), Gβ3 (A11/A'11), ABC transporter (A12/A'12), or no DNA (A2/A'2). Spots B1/B'1 through H12/H'12 contained 84 cDNA inserts isolated from clones from a cDNA library derived from a single α-gust+ TRC. Unless otherwise specified each spot contained 50 ng of a cDNA insert. Spot B3/B'3 contained lqseq91 (subsequently identified as TRPM5). (JPG 33 kb)
Supplementary Fig. 2.
Expressed sequence tag (est) matches to murine and human TRPM5 genes. For each panel the top line displays a region of ~25,000 bp containing the Trpm5/TRPM5 and adjacent Tssc1/TSSC1 genes, respectively. The second line displays the intron/exon pattern of the Trpm5/TRPM5 genes (green boxes represent exons; green lines indicate spliced out introns). The genomic sequence of the region of the TRPM5 genes from mouse (a) and human (b) were used as queries to search the mouse and human expressed sequence tag (est) databases, respectively. Matches to the est databases are shown as solid red or blue bars with red representing a higher degree of identity. Black hatched lines show gaps in the genomic sequence between contiguous sequences that had high identity matches. Matches with Trpm5/TRPM5 exons are boxed in light blue, while matches with presumed intronic sequences of Trpm5/TRPM5 are boxed in yellow. Matches to Trpm5/TRPM5 exons in the est database were few and for mouse (a) only found in embryonic or in a few cases neonatal tissues (see list of est hits below), suggesting that Trpm5/TRPM5 expression is not widespread. In contrast, the adjacent Tssc1/TSSC1 gene has many more "hits" in the est database than does Trpm5/TRPM5. There were, however, many hits from many different tissues for the Trpm5/TRPM5 est corresponding to the introns, suggesting that an expressed repetitive element or remnant of an expressed pseudogene exists within this region of Trpm5/TRPM5 genomic DNA. Tissue distribution of est matches to Trpm5 exons (demarcated within blue box in a): 12 day embryo diaphragm/neck library; 15 day embryo head library; 13 day embryo stomach library; 16 day embryo head library; 13 day embryo lung library; 14 day embryo liver library; 13 day embryo forelimb library; 0 day neonate lung; 0 day neonate eye. Tissue distribution of est matches to TRPM5 exons (demarcated within blue box in b): CGAP library, Clontech pancreas library, colon library. (GIF 32 kb)
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Pérez, C., Huang, L., Rong, M. et al. A transient receptor potential channel expressed in taste receptor cells. Nat Neurosci 5, 1169–1176 (2002). https://doi.org/10.1038/nn952
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DOI: https://doi.org/10.1038/nn952
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