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PI(4,5)P2 regulates the activation and desensitization of TRPM8 channels through the TRP domain

Nature Neuroscience volume 8, pages 626–634 (2005)Cite this article

Abstract

The subjective feeling of cold is mediated by the activation of TRPM8 channels in thermoreceptive neurons by cold or by cooling agents such as menthol. Here, we demonstrate a central role for phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) in the activation of recombinant TRPM8 channels by both cold and menthol. Moreover, we show that Ca2+ influx through these channels activates a Ca2+-sensitive phospholipase C and that the subsequent depletion of PI(4,5)P2 limits channel activity, serving as a unique mechanism for desensitization of TRPM8 channels. Finally, we find that mutation of conserved positive residues in the highly conserved proximal C-terminal TRP domain of TRPM8 and two other family members, TRPM5 and TRPV5, reduces the sensitivity of the channels for PI(4,5)P2 and increases inhibition by PI(4,5)P2 depletion. These data suggest that the TRP domain of these channels may serve as a PI(4,5)P2-interacting site and that regulation by PI(4,5)P2 is a common feature of members of the TRP channel family.

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Figure 1: Activation by PI(4,5)P2 of TRPM8 channels expressed in Xenopus oocytes.
Figure 2: Cold and menthol increase the apparent affinity of TRPM8 channels for PI(4,5)P2.
Figure 3: Effect of PI(4,5)P2 depletion in the presence and absence of menthol or icilin.
Figure 4: PI(4,5)P2 is necessary and sufficient to activate TRPM8 channels.
Figure 5: Putative PI(4,5)P2-interacting residues in the TRP domain.
Figure 6: The effect of the mutation of the equivalent residue to R1008 in TRPV5 and TRPM5 channels.
Figure 7: Calcium-dependent desensitization is mediated by PI(4,5)P2 hydrolysis in mammalian cells.
Figure 8: Calcium-induced inhibition is mediated by PI(4,5)P2 hydrolysis in Xenopus oocytes.

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Acknowledgements

We thank E. Findeis and A. Pappas for preparing oocytes. The cDNA clone for the TRPM8 channel was kindly provided by D. Julius (University of California, San Francisco); the PDGF receptor clone by A. Kazlauskas (Harvard Medical School, Boston, Massachusetts); the CFP- and YFP-tagged PH domain constructs by T. Balla (National Institutes of Health, Bethesda, Maryland); the type I IP3 5-phosphatase by C. Erneux (IRIBHN, Brussels, Belgium); the clones for PLCδ1 and type I phosphatidylinositol-4-phosphate 5-kinase (PIP5K) by S. Scarlata (State University of New York at Stony Brook) and S.-L. Syng (Oregon Health & Science University, Portland); the clone for TRPV5 (CaT2) by L. Parent (University of Montreal); and the clone for TRPM5 by R. Margolskee (Mount Sinai School of Medicine, New York). This work was supported by NIH grant HL59949 to D.L. T.R. and C.M.B.L. were supported by Scientist Development Grants from the American Heart Association.

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  1. Tibor Rohács

    Present address: Department of Pharmacology and Physiology, UMDNJ, New Jersey Medical School, 185 South Orange Ave, Newark, New Jersey, 07103, USA

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  1. Department of Physiology and Biophysics, Mount Sinai School of Medicine, One Gustave L. Levy Place, Box 1218, New York, 10029, New York, USA

    Tibor Rohács, Coeli M B Lopes, Ioannis Michailidis & Diomedes E Logothetis

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Supplementary information

Supplementary Fig. 1

PI(4,5)P2 induces a shift in the voltage dependence of TRPM8 channels. (PDF 1480 kb)

Supplementary Fig. 2

Phospholipase C is required for PDGFinhibition of TrpM8 mutants. (PDF 1856 kb)

Supplementary Fig. 3

Inhibition of the endogenous Ca2+ activated Cl− current in Cl-free media. (PDF 670 kb)

Supplementary Fig. 4

Multiple sequence alignment of the Trp-domain of the mammalian Trp superfamily. (PDF 1204 kb)

Supplementary Fig. 5

Intracellular calcium concentration measurements. (PDF 1617 kb)

Supplementary Fig. 6

Model of the activation and desensitization of TRPM8 channels. (PDF 642 kb)

Supplementary Methods (PDF 62 kb)

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Rohács, T., Lopes, C., Michailidis, I. et al. PI(4,5)P2 regulates the activation and desensitization of TRPM8 channels through the TRP domain. Nat Neurosci 8, 626–634 (2005). https://doi.org/10.1038/nn1451

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