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Direct inhibition of the cold-activated TRPM8 ion channel by Gαq

Nature Cell Biology volume 14, pages 851–858 (2012)Cite this article

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Abstract

Activation of the TRPM8 ion channel in sensory nerve endings produces a sensation of pleasant coolness. Here we show that inflammatory mediators such as bradykinin and histamine inhibit TRPM8 in intact sensory nerves, but do not do so through conventional signalling pathways. The G-protein subunit Gαq instead binds to TRPM8 and when activated by a Gq-coupled receptor directly inhibits ion channel activity. Deletion of Gαq largely abolished inhibition of TRPM8, and inhibition was rescued by a Gαq chimaera whose ability to activate downstream signalling pathways was completely ablated. Activated Gαq protein, but not Gβγ, potently inhibits TRPM8 in excised patches. We conclude that Gαq pre-forms a complex with TRPM8 and inhibits activation of TRPM8, following activation of G-protein-coupled receptors, by a direct action. This signalling mechanism may underlie the abnormal cold sensation caused by inflammation.

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Figure 1: Inflammatory mediators inhibit TRPM8-dependent cold nerve fibre activity.
Figure 2: Inflammatory mediators inhibit TRPM8 independently of downstream signalling pathways.
Figure 3: Inhibition of TRPM8 by BK is membrane-delimited.
Figure 4: Activated Gαq inhibits TRPM8 independently of the PLC pathway.
Figure 5: Inflammatory mediators inhibit TRPM8 through a direct action of activated Gαq.
Figure 6: Direct interaction of TRPM8 with Gαq.
Figure 7: Activated Gαq directly inhibits TRPM8 in excised patches.

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Acknowledgements

We thank S. Offermanns (Max Planck Institute for Heart and Lung Research, Germany) for providing MEF cells, G. Hammond (Department of Pharmacology, University of Cambridge, UK) for Tubby–R332H–cYFP and PLCδ-PH–EGFP cDNA and for help with imaging analysis, O. Opaleye (Department of Biochemistry, University of Cambridge, UK) for help with protein purification, R. Hardie (Department of Physiology, Development and Neuroscience, University of Cambridge, UK) and S.B. Hladky (Department of Pharmacology, University of Cambridge, UK) for help with single-channel recording, and R. Hardie for critical reading of an earlier version of the manuscript. This work was supported by an MRC new investigator research grant (G0801387 to X.Z.), a BBSRC research grant (BB/F003072/1 to P.A.M.) and a grant from the Fundacion BBVA (to P.A.M., to support a BBVA visiting professorship at the Instituto de Neurociencias, Alicante, Spain).

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Authors and Affiliations

  1. Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK

    Xuming Zhang, Stephanie Mak, Lin Li & Peter A. McNaughton

  2. Instituto de Neurociencias de Alicante, Universidad Miguel Hernandez-Consejo Superior de Investigaciones Cientificas, 03550 San Juan de Alicante, Spain

    Andres Parra, Bristol Denlinger & Carlos Belmonte

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  1. Xuming Zhang
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Contributions

X.Z. came up with the hypothesis, designed and performed experiments, and analysed data except calcium imaging experiments, which were carried out by S.M., and nerve fibre recordings, which were carried out by A.P, B.D., C.B. and P.A.M.; L.L. assisted with molecular biology experiments and neuron preparation. X.Z. wrote the manuscript with input from C.B. and P.A.M. X.Z. and P.A.M. supervisedthe project.

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Correspondence to Xuming Zhang or Peter A. McNaughton.

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The authors declare no competing financial interests.

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Zhang, X., Mak, S., Li, L. et al. Direct inhibition of the cold-activated TRPM8 ion channel by Gαq. Nat Cell Biol 14, 851–858 (2012). https://doi.org/10.1038/ncb2529

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