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Activity-dependent silencing reveals functionally distinct itch-generating sensory neurons

Nature Neuroscience volume 16, pages 910–918 (2013)Cite this article

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Abstract

The peripheral terminals of primary sensory neurons detect histamine and non-histamine itch-provoking ligands through molecularly distinct transduction mechanisms. It remains unclear, however, whether these distinct pruritogens activate the same or different afferent fibers. Using a strategy of reversibly silencing specific subsets of murine pruritogen-sensitive sensory axons by targeted delivery of a charged sodium-channel blocker, we found that functional blockade of histamine itch did not affect the itch evoked by chloroquine or SLIGRL-NH2, and vice versa. Notably, blocking itch-generating fibers did not reduce pain-associated behavior. However, silencing TRPV1+ or TRPA1+ neurons allowed allyl isothiocyanate or capsaicin, respectively, to evoke itch, implying that certain peripheral afferents may normally indirectly inhibit algogens from eliciting itch. These findings support the presence of functionally distinct sets of itch-generating neurons and suggest that targeted silencing of activated sensory fibers may represent a clinically useful anti-pruritic therapeutic approach for histaminergic and non-histaminergic pruritus.

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Figure 1: Application of pruritogens leads to a QX-314–mediated blockade of sodium currents selectively in pruritogen-sensitive trigeminal ganglion neurons.
Figure 2: Co-administration of QX-314 and pruritogens inhibits subsequent pruritogen-evoked scratching.
Figure 3: Silencing pruritogen-responsive neurons does not alter behavioral responses to non-itch stimuli.
Figure 4: Distinct primary afferents mediate histaminergic itch and non-histaminergic itch.
Figure 5: Proportional representation of coincident trigeminal cell responses to low-dose chloroquine and histamine and their overlapping responsiveness with capsaicin and AITC.
Figure 6: Selective silencing of nociceptor populations differentially inhibits histamine itch and non-histamine itch.
Figure 7: TRPV1+/TRPA1+ fibers suppress itch.

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References

  1. Grundmann, S. & Stander, S. Chronic pruritus: clinics and treatment. Ann. Dermatol. 23, 1–11 (2011).

    Article  Google Scholar 

  2. Yosipovitch, G. & Papoiu, A.D. What causes itch in atopic dermatitis? Curr. Allergy Asthma Rep. 8, 306–311 (2008).

    Article  CAS  Google Scholar 

  3. Lee, M.G. et al. Agonists of the MAS-related gene (Mrgs) orphan receptors as novel mediators of mast cell-sensory nerve interactions. J. Immunol. 180, 2251–2255 (2008).

    Article  CAS  Google Scholar 

  4. Akiyama, T., Carstens, M.I. & Carstens, E. Enhanced scratching evoked by PAR-2 agonist and 5-HT but not histamine in a mouse model of chronic dry skin itch. Pain 151, 378–383 (2010).

    Article  CAS  Google Scholar 

  5. Papoiu, A.D., Tey, H.L., Coghill, R.C., Wang, H. & Yosipovitch, G. Cowhage-induced itch as an experimental model for pruritus. A comparative study with histamine-induced itch. PLoS ONE 6, e17786 (2011).

    Article  CAS  Google Scholar 

  6. Liu, Q. et al. Sensory neuron–specific GPCR Mrgprs are itch receptors mediating chloroquine-induced pruritus. Cell 139, 1353–1365 (2009).

    Article  Google Scholar 

  7. Akiyama, T., Carstens, M.I. & Carstens, E. Facial injections of pruritogens and algogens excite partly overlapping populations of primary and second-order trigeminal neurons in mice. J. Neurophysiol. 104, 2442–2450 (2010).

    Article  CAS  Google Scholar 

  8. Patel, K.N. & Dong, X. Itch: cells, molecules and circuits. ACS Chem. Neurosci. 2, 17–25 (2011).

    Article  Google Scholar 

  9. Liu, Q. et al. The distinct roles of two GPCRs, MrgprC11 and PAR2, in ich and hyperalgesia. Sci. Signal. 4, 45 (2011).

    Google Scholar 

  10. Han, S.-K., Mancino, V. & Simon, M.I. Phospholipase Cbeta 3 mediates the scratching response activated by the histamine H1 receptor on C-fiber nociceptive neurons. Neuron 52, 691–703 (2006).

    Article  CAS  Google Scholar 

  11. Imamachi, N. et al. TRPV1-expressing primary afferents generate behavioral responses to pruritogens via multiple mechanisms. Proc. Natl. Acad. Sci. USA 106, 11330–11335 (2009).

    Article  CAS  Google Scholar 

  12. Wilson, S.R. et al. TRPA1 is required for histamine-independent, Mas-related G protein–coupled receptor–mediated itch. Nat. Neurosci. 14, 595–602 (2011).

    Article  CAS  Google Scholar 

  13. Akiyama, T., Carstens, M.I. & Carstens, E. Enhanced responses of lumbar superficial dorsal horn neurons to intradermal PAR-2 agonist but not histamine in a mouse hindpaw dry skin itch model. J. Neurophysiol. 105, 2811–2817 (2011).

    Article  CAS  Google Scholar 

  14. Shim, W.-S. et al. TRPV1 mediates histamine-induced itching via the activation of phospholipase A2 and 12-lipoxygenase. J. Neurosci. 27, 2331–2337 (2007).

    Article  CAS  Google Scholar 

  15. Han, L. et al. A subpopulation of nociceptors specifically linked to itch. Nat. Neurosci. 16, 174–182 (2013).

    Article  CAS  Google Scholar 

  16. Namer, B. et al. Separate peripheral pathways for pruritus in man. J. Neurophysiol. 100, 2062–2069 (2008).

    Article  Google Scholar 

  17. Liu, Q. et al. Mechanisms of itch evoked by beta-alanine. J. Neurosci. 32, 14532–14537 (2012).

    Article  CAS  Google Scholar 

  18. Akiyama, T. et al. Cross-sensitization of histamine-independent itch in mouse primary sensory neurons. Neuroscience 226, 305–312 (2012).

    Article  CAS  Google Scholar 

  19. Binshtok, A.M., Bean, B.P. & Woolf, C.J. Inhibition of nociceptors by TRPV1-mediated entry of impermeant sodium channel blockers. Nature 449, 607–610 (2007).

    Article  CAS  Google Scholar 

  20. Brenneis, C. et al. Phenotyping the function of TRPV1-expressing sensory neurons by targeted axonal silencing. J. Neurosci. 33, 315–326 (2013).

    Article  CAS  Google Scholar 

  21. Kim, H.Y. et al. Selectively targeting pain in the trigeminal system. Pain 150, 29–40 (2010).

    Article  Google Scholar 

  22. Puopolo, M. et al. Permeation and block of TRPV1 channels by the cationic lidocaine derivative QX-314. J. Neurophysiol. 109, 1704–1723 (2013).

    Article  CAS  Google Scholar 

  23. Binshtok, A.M. et al. Coapplication of lidocaine and the permanently charged sodium channel blocker QX-314 produces a long-lasting nociceptive blockade in rodents. Anesthesiology 111, 127–137 (2009).

    Article  CAS  Google Scholar 

  24. Roberson, D.P., Binshtok, A.M., Blasl, F., Bean, B.P. & Woolf, C.J. Targeting of sodium channel blockers into nociceptors to produce long-duration analgesia: a systematic study and review. Br. J. Pharmacol. 164, 48–58 (2011).

    Article  CAS  Google Scholar 

  25. Chen, J. et al. Pore dilation occurs in TRPA1 but not in TRPM8 channels. Mol. Pain 5, 3 (2009).

    PubMed  PubMed Central  Google Scholar 

  26. Shimada, S.G. & LaMotte, R.H. Behavioral differentiation between itch and pain in mouse. Pain 139, 681–687 (2008).

    Article  Google Scholar 

  27. Brenner, D.S., Golden, J.P. & Gereau, R.W. A novel behavioral assay for measuring cold sensation in mice. PLoS ONE 7, e39765 (2012).

    Article  CAS  Google Scholar 

  28. Ogle, M.E., Gu, X., Espinera, A.R. & Wei, L. Inhibition of prolyl hydroxylases by dimethyloxaloylglycine after stroke reduces ischemic brain injury and requires hypoxia inducible factor-1alpha. Neurobiol. Dis. 45, 733–742 (2012).

    Article  CAS  Google Scholar 

  29. Petersen, L.J. Quantitative measurement of extracellular histamine concentrations in intact human skin in vivo by the microdialysis technique: methodological aspects. Allergy 52, 547–555 (1997).

    Article  CAS  Google Scholar 

  30. Khalil, I.F. et al. Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations. Malar. J. 10, 249 (2011).

    Article  CAS  Google Scholar 

  31. Akiyama, T., Carstens, M.I. & Carstens, E. Differential itch- and pain-related behavioral responses and micro-opoid modulation in mice. Acta Derm. Venereol. 90, 575–581 (2010).

    Article  CAS  Google Scholar 

  32. Ross, S.E. et al. Loss of inhibitory interneurons in the dorsal spinal cord and elevated itch in Bhlhb5 mutant mice. Neuron 65, 886–898 (2010).

    Article  CAS  Google Scholar 

  33. Ohsawa, Y. & Hirasawa, N. The antagonism of histamine H1 and H4 receptors ameliorates chronic allergic dermatitis via anti-pruritic and anti-inflammatory effects in NC/Nga mice. Allergy 67, 1014–1022 (2012).

    Article  CAS  Google Scholar 

  34. Liu, T. & Ji, R.R. Oxidative stress induces itch via activation of transient receptor potential subtype ankyrin 1 in mice. Neurosci. Bull. 28, 145–154 (2012).

    Article  Google Scholar 

  35. Sun, Y.-G. et al. Cellular basis of itch sensation. Science 325, 1531–1534 (2009).

    Article  CAS  Google Scholar 

  36. Schmelz, M. et al. Chemical response pattern of different classes of C-nociceptors to pruritogens and algogens. J. Neurophysiol. 89, 2441–2448 (2003).

    Article  CAS  Google Scholar 

  37. Namer, B. & Reeh, P. Scratching an itch. Nat. Neurosci. 16, 117–118 (2013).

    Article  CAS  Google Scholar 

  38. Sun, Y.G. & Chen, Z.F. A gastrin-releasing peptide receptor mediates the itch sensation in the spinal cord. Nature 448, 700–703 (2007).

    Article  CAS  Google Scholar 

  39. Rudick, C.N., Bryce, P.J., Guichelaar, L.A., Berry, R.E. & Klumpp, D.J. Mast cell–derived histamine mediates cystitis pain. PLoS ONE 3, e2096 (2008).

    Article  Google Scholar 

  40. Sikand, P., Shimada, S.G., Green, B.G. & LaMotte, R.H. Similar itch and nociceptive sensations evoked by punctate cutaneous application of capsaicin, histamine and cowhage. Pain 144, 66–75 (2009).

    Article  CAS  Google Scholar 

  41. Koppert, W., Reeh, P.W. & Handwerker, H.O. Conditioning of histamine by bradykinin alters responses of rat nociceptor and human itch sensation. Neurosci. Lett. 152, 117–120 (1993).

    Article  CAS  Google Scholar 

  42. Liu, Y. et al. VGLUT2-dependent glutamate release from nociceptors is required to sense pain and suppress itch. Neuron 68, 543–556 (2010).

    Article  CAS  Google Scholar 

  43. Leffler, A. et al. The vanilloid receptor TRPV1 is activated and sensitized by local anesthetics in rodent sensory neurons. J. Clin. Invest. 118, 763–776 (2008).

    PubMed  PubMed Central  Google Scholar 

  44. Malin, S.A., Davis, B.M. & Molliver, D.C. Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity. Nat. Protoc. 2, 152–160 (2007).

    Article  CAS  Google Scholar 

  45. Chaplan, S.R., Bach, F.W., Pogrel, J.W., Chung, J.M. & Yaksh, T.L. Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53, 55–63 (1994).

    Article  CAS  Google Scholar 

  46. Hargreaves, K., Dubner, R., Brown, F., Flores, C. & Joris, J. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain 32, 77–88 (1988).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank N. Ghasemlou and S. Ross for discussion and O. Viramontes for technical assistance. This study was supported by the US National Institutes of Health (NS072040 to B.P.B., Q.M., B.D. and C.J.W.; NS047710 to B.D. and Q.M.). J.M.S. is a recipient of a Ruth L Kirschstein National Research Service Award from the National Institute of Dental and Craniofacial Research.

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Author notes

  1. Sagi Gudes and Jared M Sprague: These authors contributed equally to this work.

Authors and Affiliations

  1. FM Kirby Neurobiology Center and Department of Neurology, Children's Hospital, Boston, Massachusetts, USA

    David P Roberson, Jared M Sprague, Haley A W Patoski, Victoria K Robson, Seog Bae Oh & Clifford J Woolf

  2. Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA

    David P Roberson, Bo Duan, Bruce P Bean, Qiufu Ma & Clifford J Woolf

  3. Department of Medical Neurobiology, Institute for Medical Research Israel-Canada, The Hebrew University Faculty of Medicine, Jerusalem, Israel

    Sagi Gudes, Felix Blasl & Alexander M Binshtok

  4. The Edmond and Lily Safra Center for Brain Sciences, The Hebrew University, Jerusalem, Israel

    Sagi Gudes & Alexander M Binshtok

  5. Biological Sciences of Dental Medicine, Harvard School of Dental Medicine, Boston, Massachusetts, USA

    Jared M Sprague

  6. Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Bo Duan & Qiufu Ma

  7. Dental Research Institute and Department of Neurobiology and Physiology, National Research Laboratory for Pain, School of Dentistry, Seoul National University, Seoul, Republic of Korea

    Seog Bae Oh

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Contributions

D.P.R. conceived, designed and conducted the behavioral experiments, analyzed behavioral data, and wrote the manuscript. S.G. performed the combined calcium-imaging and electrophysiology experiments. J.M.S. carried out the calcium-imaging experiments. H.A.W.P. and V.K.R. conducted behavioral experiments and analyzed behavioral data. F.B. contributed to the combined calcium-imaging and electrophysiology experiments. B.D. contributed to behavioral experiments and interpretation of behavioral data. S.B.O. gathered behavioral pilot data and gave manuscript advice. B.P.B. conceived the silencing strategy, gave technical and conceptual advice, and edited the manuscript. Q.M. provided critical analysis and interpretation of behavioral data, designed behavioral experiments, and contributed to critical revision of the manuscript. A.M.B. conceived the silencing strategy, conceived and designed behavioral experiments and the combined calcium-imaging and electrophysiology experiments, gathered behavioral pilot data, supervised the project, and wrote the manuscript. C.J.W. conceived the silencing strategy, supervised the project and wrote the manuscript.

Corresponding authors

Correspondence to Alexander M Binshtok or Clifford J Woolf.

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Competing interests

This research was funded, in part, by a research grant from Endo Pharmaceuticals, who have licensed the technology invented by B.P.B. and C.J.W

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Roberson, D., Gudes, S., Sprague, J. et al. Activity-dependent silencing reveals functionally distinct itch-generating sensory neurons. Nat Neurosci 16, 910–918 (2013). https://doi.org/10.1038/nn.3404

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