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Activation of neuronal P2X7 receptor–pannexin-1 mediates death of enteric neurons during colitis

Nature Medicine volume 18, pages 600–604 (2012)Cite this article

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Abstract

Inflammatory bowel diseases (IBDs) are chronic relapsing and remitting conditions associated with long-term gut dysfunction resulting from alterations to the enteric nervous system and a loss of enteric neurons1,2. The mechanisms underlying inflammation-induced enteric neuron death are unknown. Here using in vivo models of experimental colitis we report that inflammation causes enteric neuron death by activating a neuronal signaling complex composed of P2X7 receptors (P2X7Rs), pannexin-1 (Panx1) channels, the Asc adaptor protein and caspases. Inhibition of P2X7R, Panx1, Asc or caspase activity prevented inflammation-induced neuron cell death. Preservation of enteric neurons by inhibiting Panx1 in vivo prevented the onset of inflammation-induced colonic motor dysfunction. Panx1 expression was reduced in Crohn's disease but not ulcerative colitis. We conclude that activation of neuronal Panx1 underlies neuron death and the subsequent development of abnormal gut motility in IBD. Targeting Panx1 represents a new neuroprotective strategy to ameliorate the progression of IBD-associated dysmotility.

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Figure 1: P2X7R activation is necessary and sufficient for enteric neuron death.
Figure 2: Inflammation-induced enteric neuron death requires Panx1 and Asc but not Nlrp3.
Figure 3: Inhibiting neuronal Panx1 during colitis protects against post-inflammation deficits in inhibitory neuromuscular transmission.
Figure 4: Panx1 inhibition is neuroprotective in a recurrent model of colitis, and human enteric neuron Panx1 expression changes are observed in Crohn's disease.

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References

  1. Mawe, G.M., Strong, D.S. & Sharkey, K.A. Plasticity of enteric nerve functions in the inflamed and postinflamed gut. Neurogastroenterol. Motil. 21, 481–491 (2009).

    Article  CAS  Google Scholar 

  2. De Giorgio, R. et al. Inflammatory neuropathies of the enteric nervous system. Gastroenterology 126, 1872–1883 (2004).

    Article  Google Scholar 

  3. Wynn, G., Ma, B., Ruan, H.Z. & Burnstock, G. Purinergic component of mechanosensory transduction is increased in a rat model of colitis. Am. J. Physiol. Gastrointest. Liver Physiol. 287, G647–G657 (2004).

    Article  CAS  Google Scholar 

  4. Lomax, A.E., Mawe, G.M. & Sharkey, K.A. Synaptic facilitation and enhanced neuronal excitability in the submucosal plexus during experimental colitis in guinea-pig. J. Physiol. (Lond.) 564, 863–875 (2005).

    Article  CAS  Google Scholar 

  5. Friedman, D.J. et al. From the Cover: CD39 deletion exacerbates experimental murine colitis and human polymorphisms increase susceptibility to inflammatory bowel disease. Proc. Natl. Acad. Sci. USA 106, 16788–16793 (2009).

    Article  CAS  Google Scholar 

  6. Rybaczyk, L. et al. New bioinformatics approach to analyze gene expressions and signaling pathways reveals unique purine gene dysregulation profiles that distinguish between CD and UC. Inflamm. Bowel Dis. 15, 971–984 (2009).

    Article  Google Scholar 

  7. Yiangou, Y. et al. ATP-gated ion channel P2X3 is increased in human inflammatory bowel disease. Neurogastroenterol. Motil. 13, 365–369 (2001).

    Article  CAS  Google Scholar 

  8. Guzman, J. et al. ADOA3R as a therapeutic target in experimental colitis: proof by validated high-density oligonucleotide microarray analysis. Inflamm. Bowel Dis. 12, 766–789 (2006).

    Article  Google Scholar 

  9. Atarashi, K. et al. ATP drives lamina propria TH17 cell differentiation. Nature 455, 808–812 (2008).

    Article  CAS  Google Scholar 

  10. Wang, X. et al. P2X7 receptor inhibition improves recovery after spinal cord injury. Nat. Med. 10, 821–827 (2004).

    Article  CAS  Google Scholar 

  11. Lazarowski, E.R., Boucher, R.C. & Harden, T.K. Constitutive release of ATP and evidence for major contribution of ecto-nucleotide pyrophosphatase and nucleoside diphosphokinase to extracellular nucleotide concentrations. J. Biol. Chem. 275, 31061–31068 (2000).

    Article  CAS  Google Scholar 

  12. Sperlágh, B., Vizi, E.S., Wirkner, K. & Illes, P. P2X7 receptors in the nervous system. Prog. Neurobiol. 78, 327–346 (2006).

    Article  Google Scholar 

  13. Cavaliere, F., Amadio, S., Sancesario, G., Bernardi, G. & Volonte, C. Synaptic P2X7 and oxygen/glucose deprivation in organotypic hippocampal cultures. J. Cereb. Blood Flow Metab. 24, 392–398 (2004).

    Article  CAS  Google Scholar 

  14. Hu, H. et al. Stimulation of the P2X7 receptor kills rat retinal ganglion cells in vivo. Exp. Eye Res. 91, 425–432 (2010).

    Article  CAS  Google Scholar 

  15. Zhang, X., Zhang, M., Laties, A.M. & Mitchell, C.H. Stimulation of P2X7 receptors elevates Ca2+ and kills retinal ganglion cells. Invest. Ophthalmol. Vis. Sci. 46, 2183–2191 (2005).

    Article  Google Scholar 

  16. Linden, D.R. et al. Indiscriminate loss of myenteric neurones in the TNBS-inflamed guinea-pig distal colon. Neurogastroenterol. Motil. 17, 751–760 (2005).

    Article  CAS  Google Scholar 

  17. Locovei, S., Scemes, E., Qiu, F., Spray, D.C. & Dahl, G. Pannexin1 is part of the pore forming unit of the P2X7 receptor death complex. FEBS Lett. 581, 483–488 (2007).

    Article  CAS  Google Scholar 

  18. Seminario-Vidal, L. et al. Rho signaling regulates pannexin 1–mediated ATP release from airway epithelia. J. Biol. Chem. 286, 26277–26286 (2011).

    Article  CAS  Google Scholar 

  19. Thompson, R.J. et al. Activation of pannexin-1 hemichannels augments aberrant bursting in the hippocampus. Science 322, 1555–1559 (2008).

    Article  CAS  Google Scholar 

  20. Ferrari, D. et al. P2Z purinoreceptor ligation induces activation of caspases with distinct roles in apoptotic and necrotic alterations of cell death. FEBS Lett. 447, 71–75 (1999).

    Article  CAS  Google Scholar 

  21. Ferrari, D. et al. Extracellular ATP triggers IL-1β release by activating the purinergic P2Z receptor of human macrophages. J. Immunol. 159, 1451–1458 (1997).

    CAS  Google Scholar 

  22. Silverman, W., Locovei, S. & Dahl, G. Probenecid, a gout remedy, inhibits pannexin 1 channels. Am. J. Physiol. Cell Physiol. 295, C761–C767 (2008).

    Article  CAS  Google Scholar 

  23. Silverman, W.R. et al. The pannexin 1 channel activates the inflammasome in neurons and astrocytes. J. Biol. Chem. 284, 18143–18151 (2009).

    Article  CAS  Google Scholar 

  24. Chekeni, F.B. et al. Pannexin 1 channels mediate 'find-me' signal release and membrane permeability during apoptosis. Nature 467, 863–867 (2010).

    Article  CAS  Google Scholar 

  25. Gulbransen, B.D. & Sharkey, K.A. Purinergic neuron-to-glia signaling in the enteric nervous system. Gastroenterology 136, 1349–1358 (2009).

    Article  CAS  Google Scholar 

  26. Gomes, P. et al. ATP-dependent paracrine communication between enteric neurons and glia in a primary cell culture derived from embryonic mice. Neurogastroenterol. Motil. 21, 870–e62 (2009).

    Article  CAS  Google Scholar 

  27. Collins, S.M. The immunomodulation of enteric neuromuscular function: implications for motility and inflammatory disorders. Gastroenterology 111, 1683–1699 (1996).

    Article  CAS  Google Scholar 

  28. Vasina, V. et al. Enteric neuroplasticity evoked by inflammation. Auton. Neurosci. 126–127, 264–272 (2006).

    Article  Google Scholar 

  29. Krauter, E.M. et al. Changes in colonic motility and the electrophysiological properties of myenteric neurons persist following recovery from trinitrobenzene sulfonic acid colitis in the guinea pig. Neurogastroenterol. Motil. 19, 990–1000 (2007).

    CAS  PubMed  Google Scholar 

  30. Bossone, C., Hosseini, J.M., Pineiro-Carrero, V. & Shea-Donohue, T. Alterations in spontaneous contractions in vitro after repeated inflammation of rat distal colon. Am. J. Physiol. Gastrointest. Liver Physiol. 280, G949–G957 (2001).

    Article  CAS  Google Scholar 

  31. Mizuta, Y., Isomoto, H. & Takahashi, T. Impaired nitrergic innervation in rat colitis induced by dextran sulfate sodium. Gastroenterology 118, 714–723 (2000).

    Article  CAS  Google Scholar 

  32. Depoortere, I., Thijs, T. & Peeters, T.L. Generalized loss of inhibitory innervation reverses serotonergic inhibition into excitation in a rabbit model of TNBS-colitis. Br. J. Pharmacol. 135, 2011–2019 (2002).

    Article  CAS  Google Scholar 

  33. Strong, D.S. et al. Purinergic neuromuscular transmission is selectively attenuated in ulcerated regions of inflamed guinea pig distal colon. J. Physiol. (Lond.) 588, 847–859 (2010).

    Article  CAS  Google Scholar 

  34. McCafferty, D.M. et al. Spontaneously developing chronic colitis in IL-10/iNOS double-deficient mice. Am. J. Physiol. Gastrointest. Liver Physiol. 279, G90–G99 (2000).

    Article  CAS  Google Scholar 

  35. Wirtz, S., Neufert, C., Weigmann, B. & Neurath, M.F. Chemically induced mouse models of intestinal inflammation. Nat. Protoc. 2, 541–546 (2007).

    Article  CAS  Google Scholar 

  36. Storr, M.A. et al. Activation of the cannabinoid 2 receptor (CB2) protects against experimental colitis. Inflamm. Bowel Dis. 15, 1678–1685 (2009).

    Article  Google Scholar 

  37. Gulbransen, B.D., Bains, J.S. & Sharkey, K.A. Enteric glia are targets of the sympathetic innervation of the myenteric plexus in the guinea pig distal colon. J. Neurosci. 30, 6801–6809 (2010).

    Article  CAS  Google Scholar 

  38. Nasser, Y. et al. Role of enteric glia in intestinal physiology: effects of the gliotoxin fluorocitrate on motor and secretory function. Am. J. Physiol. Gastrointest. Liver Physiol. 291, G912–G927 (2006).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Canadian Institutes of Health Research (CIHR, to K.A.S., R.J.T. and D.M.M.), the Crohn's & Colitis Foundation of Canada (CCFC to D.M.M.) and US National Institutes of Health grant DK62267 (to G.M.M.). Some of the equipment used in the study was provided by funds from the Canadian Foundation for Innovation and the Alberta Science and Research Authority. B.D.G. holds fellowships from the Canadian Association of Gastroenterology/CIHR and Alberta Innovates-Health Solutions (AI-HS)/CCFC. K.A.S. is an AI-HS Medical Scientist and holds the CCFC Chair in IBD Research at the University of Calgary. R.J.T. is an AI-HS Scholar. P.L.B. is supported by AI-HS, CIHR and CCFC. J.A.M. is an AI-HS Senior Scholar and Canada Research Chair. D.M.M. is an AI-HS Scientist and holds a Canada Research Chair (Tier 1). S.A.H. holds fellowships from CIHR and AI-HS. We thank C. MacNaughton, W. Ho and A. Wang for technical support, D.M. McCafferty (University of Calgary) for providing Il10−/− mice, V. Dixit (Genentech) and J. Tschopp (University of Lausanne) for providing male Pycard−/− and Nlrp3–/– mice, respectively, N. Hyman (University of Vermont), O. Bathe (University of Calgary) and the IBD Tissue Bank for providing human tissue samples and J. Bains for reviewing and commenting on the manuscript.

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Authors and Affiliations

  1. Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada

    Brian D Gulbransen, Mohammad Bashashati, Roger J Thompson & Keith A Sharkey

  2. Department of Physiology and Pharmacology, University of Calgary, Calgary, Alberta, Canada

    Brian D Gulbransen, Mohammad Bashashati, Derek M McKay & Keith A Sharkey

  3. Snyder Institute of Infection, Immunity and Inflammation, University of Calgary, Calgary, Alberta, Canada

    Brian D Gulbransen, Mohammad Bashashati, Simon A Hirota, Justin A MacDonald, Daniel A Muruve, Derek M McKay, Paul L Beck & Keith A Sharkey

  4. Department of Medicine, University of Calgary, Calgary, Alberta, Canada

    Simon A Hirota, Daniel A Muruve & Paul L Beck

  5. Libin Cardiovascular Institute, University of Calgary, Calgary, Alberta, Canada

    Simon A Hirota & Justin A MacDonald

  6. Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada

    Simon A Hirota & Justin A MacDonald

  7. Department of Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada

    Xianyong Gui

  8. Department of Anatomy & Neurobiology, University of Vermont College of Medicine, Burlington, Vermont, USA

    Jane A Roberts & Gary M Mawe

  9. Department of Cell Biology and Anatomy, University of Calgary, Calgary, Alberta, Canada

    Roger J Thompson

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Contributions

Overall project design and hypotheses were developed by B.D.G. under the supervision of R.J.T. and K.A.S. B.D.G. coordinated the project, conducted all experiments unless otherwise noted, analyzed the data, prepared figures and wrote the manuscript. M.B. carried out the colon contractility experiments. P.L.B., X.G., S.A.H., J.A.R., J.A.M., D.A.M., D.M.M. and G.M.M. contributed to experimental design and prepared and provided mice, tissues and reagents. All authors participated in revising the manuscript and agreed to the final version. R.J.T. and K.A.S. supervised the overall project.

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Correspondence to Brian D Gulbransen or Keith A Sharkey.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Table 1 and Supplementary Methods (PDF 83820 kb)

Supplementary Video 1

A myenteric ganglia containing enteric neurons and glia (loaded with the Ca2+ indicator Fluo4) is challenged with the P2X7R agonist BzATP. Note how the response begins in neurons, followed closely by glia. (AVI 1205 kb)

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Gulbransen, B., Bashashati, M., Hirota, S. et al. Activation of neuronal P2X7 receptor–pannexin-1 mediates death of enteric neurons during colitis. Nat Med 18, 600–604 (2012). https://doi.org/10.1038/nm.2679

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