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.

Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition

Abstract

Tissue injury generates endogenous factors that heighten our sense of pain by increasing the response of sensory nerve endings to noxious stimuli1,2. Bradykinin and nerve growth factor (NGF) are two such pro-algesic agents that activate G-protein-coupled (BK2) and tyrosine kinase (TrkA) receptors, respectively, to stimulate phospholipase C (PLC) signalling pathways in primary afferent neurons3,4. How these actions produce sensitization to physical or chemical stimuli has not been elucidated at the molecular level. Here, we show that bradykinin- or NGF-mediated potentiation of thermal sensitivity in vivo requires expression of VR1, a heat-activated ion channel on sensory neurons. Diminution of plasma membrane phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) levels through antibody sequestration or PLC-mediated hydrolysis mimics the potentiating effects of bradykinin or NGF at the cellular level. Moreover, recruitment of PLC-γ to TrkA is essential for NGF-mediated potentiation of channel activity, and biochemical studies suggest that VR1 associates with this complex. These studies delineate a biochemical mechanism through which bradykinin and NGF produce hypersensitivity and might explain how the activation of PLC signalling systems regulates other members of the TRP channel family.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1: VR1 is essential for the development of bradykinin- or NGF-evoked thermal hypersensitivity in vivo.
Figure 2: Bradykinin (BK) and nerve growth factor (NGF) sensitize VR1 in heterologous expression systems.
Figure 3: Phospholipase C is involved in nerve growth factor (NGF) modulation of VR1 function.
Figure 4: Phosphatidyl-4,5-inositol bisphosphate (PtdIns(4,5)P2) antibody application mimics modulation of VR1 by bradykinin (BK) or nerve growth factor (NGF).
Figure 5: VR1, TrkA and phospholipase Cγ (PLC-γ) form a signalling complex.

References

  1. McMahon, S. B. & Bennett, D. L. H. in Textbook of Pain (eds Wall, P. D. & Melzack, R.) 105–128 (Harcourt, London, 1999).

    Google Scholar 

  2. Bevan, S. in Textbook of Pain (eds Wall, P. D. & Melzack, R.) 85–103 (Harcourt, London, 1999).

    Google Scholar 

  3. Ganju, P., O'Bryan, J. P., Der, C., Winter, J. & James, I. F. Differential regulation of SHC proteins by nerve growth factor in sensory neurons and PC12 cells. Eur. J. Neurosci. 10, 1995–2008 (1998).

    Article  CAS  Google Scholar 

  4. Burgess, G. M., Mullaney, I., McNeill, M., Dunn, P. M. & Rang, H. P. Second messengers involved in the mechanism of action of bradykinin in sensory neurons in culture. J. Neurosci. 9, 3314–3325 (1989).

    Article  CAS  Google Scholar 

  5. Woolf, C. J. & Salter, M. W. Neuronal plasticity: increasing the gain in pain. Science 288, 1765–1769 (2000).

    Article  ADS  CAS  Google Scholar 

  6. Shu, X. Q. & Mendell, L. M. Neurotrophins and hyperalgesia. Proc. Natl Acad. Sci. USA 96, 7693–7696 (1999).

    Article  ADS  CAS  Google Scholar 

  7. Nicholas, R. S., Winter, J., Wren, P., Bergmann, R. & Woolf, C. J. Peripheral inflammation increases the capsaicin sensitivity of dorsal root ganglion neurons in a nerve growth factor-dependent manner. Neuroscience 91, 1425–1433 (1999).

    Article  CAS  Google Scholar 

  8. Koltzenburg, M., Bennett, D. L., Shelton, D. L. & McMahon, S. B. Neutralization of endogenous NGF prevents the sensitization of nociceptors supplying inflamed skin. Eur. J. Neurosci. 11, 1698–1704 (1999).

    Article  CAS  Google Scholar 

  9. Woolf, C. J. Phenotypic modification of primary sensory neurons: the role of nerve growth factor in the production of persistent pain. Phil. Trans. R. Soc. Lond. B 351, 441–448 (1996).

    Article  ADS  CAS  Google Scholar 

  10. Koltzenburg, M. The changing sensitivity in the life of the nociceptor. Pain 6 (Suppl.), S93–S102 (1999).

    Article  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  12. Tominaga, M. et al. The cloned capsaicin receptor integrates multiple pain-producing stimuli. Neuron 21, 531–543 (1998).

    Article  CAS  Google Scholar 

  13. Caterina, M. J. et al. Impaired nociception and pain sensation in mice lacking the capsaicin receptor. Science 288, 306–313 (2000).

    Article  ADS  CAS  Google Scholar 

  14. Davis, J. B. et al. Vanilloid receptor-1 is essential for inflammatory thermal hyperalgesia. Nature 405, 183–187 (2000).

    Article  ADS  CAS  Google Scholar 

  15. Jordt, S. E., Tominaga, M. & Julius, D. Acid potentiation of the capsaicin receptor determined by a key extracellular site. Proc. Natl Acad. Sci. USA 97, 8134–8139 (2000).

    Article  ADS  CAS  Google Scholar 

  16. Harteneck, C., Plant, T. D. & Schultz, G. From worm to man: three subfamilies of TRP channels. Trends Neurosci. 23, 159–166 (2000).

    Article  CAS  Google Scholar 

  17. Shu, X. & Mendell, L. M. Nerve growth factor acutely sensitizes the response of adult rat sensory neurons to capsaicin. Neurosci. Lett. 274, 159–162 (1999).

    Article  CAS  Google Scholar 

  18. Bergmann, I., Reiter, R., Toyka, K. V. & Koltzenburg, M. Nerve growth factor evokes hyperalgesia in mice lacking the low-affinity neurotrophin receptor p75. Neurosci. Lett. 255, 87–90 (1998).

    Article  CAS  Google Scholar 

  19. Stephens, R. M. et al. Trk receptors use redundant signal transduction pathways involving SHC and PLC-γ1 to mediate NGF responses. Neuron 12, 691–705 (1994).

    Article  CAS  Google Scholar 

  20. Cesare, P., Dekker, L. V., Sardini, A., Parker, P. J. & McNaughton, P. A. Specific involvement of PKC-epsilon in sensitization of the neuronal response to painful heat. Neuron 23, 617–624 (1999).

    Article  CAS  Google Scholar 

  21. Premkumar, L. S. & Ahern, G. P. Induction of vanilloid receptor channel activity by protein kinase C. Nature 408, 985–990 (2000).

    Article  ADS  CAS  Google Scholar 

  22. Womack, K. B. et al. Do phosphatidylinositides modulate vertebrate phototransduction? J. Neurosci. 20, 2792–2799 (2000).

    Article  CAS  Google Scholar 

  23. Huang, C. L., Feng, S. & Hilgemann, D. W. Direct activation of inward rectifier potassium channels by PIP2 and its stabilization by Gβγ. Nature 391, 803–806 (1998).

    Article  ADS  CAS  Google Scholar 

  24. Zhang, H., He, C., Yan, X., Mirshahi, T. & Logothetis, D. E. Activation of inwardly rectifying K+ channels by distinct PtdIns(4,5)P2 interactions. Nature Cell Biol. 1, 183–188 (1999).

    Article  CAS  Google Scholar 

  25. Li, H. S., Xu, X. Z. & Montell, C. Activation of a TRPC3-dependent cation current through the neurotrophin BDNF. Neuron 24, 261–273 (1999).

    Article  CAS  Google Scholar 

  26. Estacion, M., Sinkins, W. G. & Schilling, W. P. Regulation of Drosophila transient receptor potential-like (TrpL) channels by phospholipase C-dependent mechanisms. J. Physiol. 530, 1–19 (2001).

    Article  CAS  Google Scholar 

  27. Zygmunt, P. M. et al. Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400, 452–457 (1999).

    Article  ADS  CAS  Google Scholar 

  28. Hofmann, T. et al. Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol. Nature 397, 259–263 (1999).

    Article  ADS  CAS  Google Scholar 

  29. Chevesich, J., Kreuz, A. J. & Montell, C. Requirement for the PDZ domain protein, INAD, for localization of the TRP store-operated channel to a signaling complex. Neuron 18, 95–105 (1997).

    Article  CAS  Google Scholar 

  30. Scott, K. & Zuker, C. S. Assembly of the Drosophila phototransduction cascade into a signalling complex shapes elementary responses. Nature 395, 805–808 (1998).

    Article  ADS  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to W. Neuhausser for providing dissociated DRG neurons, W. Mobley for 2.5 S NGF, D. Shelton for advice regarding NGF administration and J. Poblete and K. Simpson for technical assistance. We thank members of our laboratories for many helpful suggestions and constructive criticism. This work was supported by a NSF predoctoral fellowship (E.P.), NIH predoctoral neuroscience training grant (S.S.), a German Academy of Natural Scientists Leopoldina postdoctoral fellowship (S.-E.J.) and by the NIH (M.V.C., A.I.B. and D.J.).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to David Julius.

Supplementary information

Supplementary Figure 1

(A) In transfected HEK cells, BK (20 nM) elicits a strongly rectifying VR1 basal current that is inhibited by 5 mM capsazepine, as shown by the holding current trace (left) and the ramp traces (right). (B) PIP2 Ab-induced VR1 basal current (-60 mV) is suppressed by 3 mM capsazepine; representative ramp traces from specified time points (* and **) are shown at right. (PDF 26 kb)

Supplementary Figure 2

[3H]-RTX binding to membranes from VR1-transfected HEK293 cells is displaced by TPA. Data are plotted as percentage of specific [3H]-RTX binding. IC50 for TPA = 256 ± 8 nM (average of four independent experiments; each point in duplicate). Note that phorbol-12,13-dibutyrate (PDBu) did not affect [3H]-RTX binding. Data were fit to the Hill equation. (DOC 33 kb)

Supplementary Figure 3

VR1 associates with wild type TrkA and TrkA mutants. HEK cells were transfected with equal amounts of VR1 and TrkA or TrkA mutant cDNAs. Immune complexes (above) were formed and analyzed as described in text. Western blots (below) of 40 mg total soluble HEK protein indicate equal expression levels of VR1 and Trks among samples. (PDF 21 kb)

Supplementary Figure 4

Effects of EGFR activation on VR1 currents. Two-electrode voltage clamp analysis was performed as described in text. Representative traces are shown above, and quantitation of 5 VR1 injected and 8 VR1/EGFR injected oocytes is shown below as SEM, p < 0.005. Note calcium activated chloride current (denoted with star) indicative of EGFR stimulation and consequent PLC activation. (PDF 26 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chuang, Hh., Prescott, E., Kong, H. et al. Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition. Nature 411, 957–962 (2001). https://doi.org/10.1038/35082088

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/35082088

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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