Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Opening of an alternative ion permeation pathway in a nociceptor TRP channel

Nature Chemical Biology volume 10, pages 188–195 (2014)Cite this article

Subjects

Abstract

Sensory neurons detect chemical stimuli through projections in the skin and mucosa, where several transient receptor potential (TRP) channels act as primary chemosensors. TRP channels are tetramers, and it is generally accepted that binding of ligands causes the opening of a single central cation-conducting pore. Contrary to this view, we here provide evidence for a second permeation pathway in the TRP channel TRPM3, which can be gated by combined application of endogenous neurosteroids and exogenous chemicals such as clotrimazole or several structurally related drugs. This alternative pathway is preserved following desensitization, blockade, mutagenesis and chemical modification of the central pore and enables massive Na+ influx at negative voltages. Opening of this alternative pathway can enhance excitation of sensory neurons and thereby exacerbate TRPM3-dependent pain. Our findings indicate that a single sensory TRP channel can encompass two distinct ionotropic chemoreceptors, which may have important ramifications for TRP channel function and pharmacology.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Activation of a new inwardly rectifying TRPM3 current component.
Figure 2: The effect of Clt is independent of CYP modulation or neurosteroid metabolism.
Figure 3: Dissection of two distinct current components in TRPM3.
Figure 4: Properties of the Clt-induced current cell-attached patches.
Figure 5: Mutations in TRPM3 differentially affect the two current components.
Figure 6: Clt potentiates TRPM3-mediated neuronal responses and pain.

Similar content being viewed by others

References

  1. Nilius, B., Owsianik, G., Voets, T. & Peters, J.A. Transient receptor potential cation channels in disease. Physiol. Rev. 87, 165–217 (2007).

    Article  CAS  Google Scholar 

  2. Hoenderop, J.G. et al. Homo- and heterotetrameric architecture of the epithelial Ca2+ channels TRPV5 and TRPV6. EMBO J. 22, 776–785 (2003).

    Article  CAS  Google Scholar 

  3. Wu, L.J., Sweet, T.B. & Clapham, D.E. International Union of Basic and Clinical Pharmacology. LXXVI. Current progress in the mammalian TRP ion channel family. Pharmacol. Rev. 62, 381–404 (2010).

    Article  CAS  Google Scholar 

  4. Yu, F.H., Yarov-Yarovoy, V., Gutman, G.A. & Catterall, W.A. Overview of molecular relationships in the voltage-gated ion channel superfamily. Pharmacol. Rev. 57, 387–395 (2005).

    Article  CAS  Google Scholar 

  5. Tombola, F., Pathak, M.M. & Isacoff, E.Y. Voltage-sensing arginines in a potassium channel permeate and occlude cation-selective pores. Neuron 45, 379–388 (2005).

    Article  CAS  Google Scholar 

  6. Sokolov, S., Scheuer, T. & Catterall, W.A. Ion permeation through a voltage-sensitive gating pore in brain sodium channels having voltage sensor mutations. Neuron 47, 183–189 (2005).

    Article  CAS  Google Scholar 

  7. Sokolov, S., Scheuer, T. & Catterall, W.A. Gating pore current in an inherited ion channelopathy. Nature 446, 76–78 (2007).

    Article  CAS  Google Scholar 

  8. Tombola, F., Pathak, M.M., Gorostiza, P. & Isacoff, E.Y. The twisted ion-permeation pathway of a resting voltage-sensing domain. Nature 445, 546–549 (2007).

    Article  CAS  Google Scholar 

  9. Wagner, T.F. et al. Transient receptor potential M3 channels are ionotropic steroid receptors in pancreatic beta cells. Nat. Cell Biol. 10, 1421–1430 (2008).

    Article  CAS  Google Scholar 

  10. Vriens, J. et al. TRPM3 is a nociceptor channel involved in the detection of noxious heat. Neuron 70, 482–494 (2011).

    Article  CAS  Google Scholar 

  11. Oberwinkler, J., Lis, A., Giehl, K.M., Flockerzi, V. & Philipp, S.E. Alternative splicing switches the divalent cation selectivity of TRPM3 channels. J. Biol. Chem. 280, 22540–22548 (2005).

    Article  CAS  Google Scholar 

  12. Wulff, H. et al. Design of a potent and selective inhibitor of the intermediate-conductance Ca2+-activated K+ channel, IKCa1: a potential immunosuppressant. Proc. Natl. Acad. Sci. USA 97, 8151–8156 (2000).

    Article  CAS  Google Scholar 

  13. Hill, K., McNulty, S. & Randall, A.D. Inhibition of TRPM2 channels by the antifungal agents clotrimazole and econazole. Naunyn Schmiedebergs Arch. Pharmacol. 370, 227–237 (2004).

    Article  CAS  Google Scholar 

  14. Meseguer, V. et al. Transient receptor potential channels in sensory neurons are targets of the antimycotic agent clotrimazole. J. Neurosci. 28, 576–586 (2008).

    Article  CAS  Google Scholar 

  15. Chung, M.K., Guler, A.D. & Caterina, M.J. TRPV1 shows dynamic ionic selectivity during agonist stimulation. Nat. Neurosci. 11, 555–564 (2008).

    Article  CAS  Google Scholar 

  16. Banke, T.G., Chaplan, S.R. & Wickenden, A.D. Dynamic changes in the TRPA1 selectivity filter lead to progressive but reversible pore dilation. Am. J. Physiol. Cell Physiol. 298, C1457–C1468 (2010).

    Article  CAS  Google Scholar 

  17. Karashima, Y. et al. Agonist-induced changes in Ca2+ permeation through the nociceptor cation channel TRPA1. Biophys. J. 98, 773–783 (2010).

    Article  CAS  Google Scholar 

  18. McNaughton-Smith, G.A. et al. Novel inhibitors of the Gardos channel for the treatment of sickle cell disease. J. Med. Chem. 51, 976–982 (2008).

    Article  CAS  Google Scholar 

  19. Jordan, V.C. Tamoxifen: a most unlikely pioneering medicine. Nat. Rev. Drug Discov. 2, 205–213 (2003).

    Article  CAS  Google Scholar 

  20. Janssens, A. & Voets, T. Ligand stoichiometry of the cold- and menthol-activated channel TRPM8. J. Physiol. (Lond.) 589, 4827–4835 (2011).

    Article  CAS  Google Scholar 

  21. Voets, T. et al. The principle of temperature-dependent gating in cold- and heat-sensitive TRP channels. Nature 430, 748–754 (2004).

    Article  CAS  Google Scholar 

  22. Voets, T., Owsianik, G., Janssens, A., Talavera, K. & Nilius, B. TRPM8 voltage sensor mutants reveal a mechanism for integrating thermal and chemical stimuli. Nat. Chem. Biol. 3, 174–182 (2007).

    Article  CAS  Google Scholar 

  23. Clapham, D.E. TRP channels as cellular sensors. Nature 426, 517–524 (2003).

    Article  CAS  Google Scholar 

  24. Caterina, M.J. et al. The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 389, 816–824 (1997).

    Article  CAS  Google Scholar 

  25. Nagata, K., Duggan, A., Kumar, G. & Garcia-Anoveros, J. Nociceptor and hair cell transducer properties of TRPA1, a channel for pain and hearing. J. Neurosci. 25, 4052–4061 (2005).

    Article  CAS  Google Scholar 

  26. McKemy, D.D., Neuhausser, W.M. & Julius, D. Identification of a cold receptor reveals a general role for TRP channels in thermosensation. Nature 416, 52–58 (2002).

    Article  CAS  Google Scholar 

  27. Grimm, C., Kraft, R., Sauerbruch, S., Schultz, G. & Harteneck, C. Molecular and functional characterization of the melastatin-related cation channel TRPM3. J. Biol. Chem. 278, 21493–21501 (2003).

    Article  CAS  Google Scholar 

  28. Havliková, H., Hill, M., Hampl, R. & Starka, L. Sex- and age-related changes in epitestosterone in relation to pregnenolone sulfate and testosterone in normal subjects. J. Clin. Endocrinol. Metab. 87, 2225–2231 (2002).

    Article  Google Scholar 

  29. Morley, J.E. et al. Potentially predictive and manipulable blood serum correlates of aging in the healthy human male: progressive decreases in bioavailable testosterone, dehydroepiandrosterone sulfate, and the ratio of insulin-like growth factor 1 to growth hormone. Proc. Natl. Acad. Sci. USA 94, 7537–7542 (1997).

    Article  CAS  Google Scholar 

  30. Virginio, C., MacKenzie, A., Rassendren, F.A., North, R.A. & Surprenant, A. Pore dilation of neuronal P2X receptor channels. Nat. Neurosci. 2, 315–321 (1999).

    Article  CAS  Google Scholar 

  31. McNally, B.A., Somasundaram, A., Yamashita, M. & Prakriya, M. Gated regulation of CRAC channel ion selectivity by STIM1. Nature 482, 241–245 (2012).

    Article  CAS  Google Scholar 

  32. Tai, K.K. & Goldstein, S.A. The conduction pore of a cardiac potassium channel. Nature 391, 605–608 (1998).

    Article  CAS  Google Scholar 

  33. Nilius, B. et al. The Ca2+-activated cation channel TRPM4 is regulated by phosphatidylinositol 4,5-biphosphate. EMBO J. 25, 467–478 (2006).

    Article  CAS  Google Scholar 

  34. Runnels, L.W., Yue, L. & Clapham, D.E. TRP-PLIK, a bifunctional protein with kinase and ion channel activities. Science 291, 1043–1047 (2001).

    Article  CAS  Google Scholar 

  35. Ciurtin, C. et al. TRPM3 channel stimulated by pregnenolone sulphate in synovial fibroblasts and negatively coupled to hyaluronan. BMC Musculoskelet. Disord. 11, 111 (2010).

    Article  Google Scholar 

  36. Czerninski, R. et al. A novel sustained-release clotrimazole varnish for local treatment of oral candidiasis. Clin. Oral Investig. 14, 71–78 (2010).

    Article  Google Scholar 

  37. Torres-Rodríguez, J.M. et al. In vitro susceptibilities of clinical yeast isolates to the new antifungal eberconazole compared with their susceptibilities to clotrimazole and ketoconazole. Antimicrob. Agents Chemother. 43, 1258–1259 (1999).

    Article  Google Scholar 

  38. Binet, O. et al. Flutrimazole 1% dermal cream in the treatment of dermatomycoses: a randomized, multicentre, double-blind, comparative clinical trial with 1% clotrimazole cream. Flutrimazole Study Group. Mycoses 37, 455–459 (1994).

    Article  CAS  Google Scholar 

  39. del Palacio, A. et al. A double-blind randomized comparative trial: eberconazole 1% cream versus clotrimazole 1% cream twice daily in Candida and dermatophyte skin infections. Mycoses 44, 173–180 (2001).

    Article  CAS  Google Scholar 

  40. Gaudioso, C., Hao, J., Martin-Eauclaire, M.F., Gabriac, M. & Delmas, P. Menthol pain relief through cumulative inactivation of voltage-gated sodium channels. Pain 153, 473–484 (2012).

    Article  CAS  Google Scholar 

  41. Erlij, D., De Smet, P. & Van Driessche, W. Effect of insulin on area and Na+ channel density of apical membrane of cultured toad kidney cells. J. Physiol. (Lond.) 481, 533–542 (1994).

    Article  CAS  Google Scholar 

  42. Karashima, Y. et al. TRPA1 acts as a cold sensor in vitro and in vivo. Proc. Natl. Acad. Sci. USA 106, 1273–1278 (2009).

    Article  CAS  Google Scholar 

  43. Kwan, K.Y. et al. TRPA1 contributes to cold, mechanical, and chemical nociception but is not essential for hair-cell transduction. Neuron 50, 277–289 (2006).

    Article  CAS  Google Scholar 

  44. Voets, T., Janssens, A., Droogmans, G. & Nilius, B. Outer pore architecture of a Ca2+-selective TRP channel. J. Biol. Chem. 279, 15223–15230 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank all members of the Laboratory of Ion Channel Research for helpful discussions. We thank the Cell Banking Facility of the Physiology Department at the Institute of Functional Genomics (Montpellier, France) for the F11 cell line. This work was supported by grants from the Belgian Federal Government (IUAP P7/13 to T.V.), the Research Foundation-Flanders (G.0565.07 and G.0825.11 to T.V. and J.V.), the Research Council of the KU Leuven (GOA 2009/07 and PF-TRPLe to T.V. and R.V.), the Planckaert-De Waele fund (to J.V.) and by a Marie Curie Intra-European Fellowship within the Seventh European Community Framework Programme (to B.I.T.).

Author information

Author notes

  1. Katharina Held, Annelies Janssens and Balázs István Tóth: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Ion Channel Research and TRP Research Platform Leuven (TRPLe), KU Leuven, Leuven, Belgium

    Joris Vriens, Katharina Held, Annelies Janssens, Balázs István Tóth, Sara Kerselaers, Bernd Nilius, Rudi Vennekens & Thomas Voets

  2. Laboratory of Obstetrics and Experimental Gynaecology, KU Leuven, Leuven, Belgium

    Joris Vriens & Katharina Held

Authors
  1. Joris Vriens
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Katharina Held
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Annelies Janssens
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Balázs István Tóth
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sara Kerselaers
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bernd Nilius
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rudi Vennekens
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Thomas Voets
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

J.V. and T.V. designed the project strategy and wrote the manuscript. J.V., K.H., A.J., B.I.T., S.K. and T.V. conducted and analyzed experiments. J.V., B.N., R.V. and T.V. supervised the project.

Corresponding authors

Correspondence to Joris Vriens or Thomas Voets.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Figures 1–13. (PDF 2227 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vriens, J., Held, K., Janssens, A. et al. Opening of an alternative ion permeation pathway in a nociceptor TRP channel. Nat Chem Biol 10, 188–195 (2014). https://doi.org/10.1038/nchembio.1428

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio.1428

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing