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Blocking microglial pannexin-1 channels alleviates morphine withdrawal in rodents

An Erratum to this article was published on 06 June 2017

This article has been updated

Abstract

Opiates are essential for treating pain, but termination of opiate therapy can cause a debilitating withdrawal syndrome in chronic users. To alleviate or avoid the aversive symptoms of withdrawal, many of these individuals continue to use opiates1,2,3,4. Withdrawal is therefore a key determinant of opiate use in dependent individuals, yet its underlying mechanisms are poorly understood and effective therapies are lacking. Here, we identify the pannexin-1 (Panx1) channel as a therapeutic target in opiate withdrawal. We show that withdrawal from morphine induces long-term synaptic facilitation in lamina I and II neurons within the rodent spinal dorsal horn, a principal site of action for opiate analgesia. Genetic ablation of Panx1 in microglia abolished the spinal synaptic facilitation and ameliorated the sequelae of morphine withdrawal. Panx1 is unique in its permeability to molecules up to 1 kDa in size and its release of ATP5,6. We show that Panx1 activation drives ATP release from microglia during morphine withdrawal and that degrading endogenous spinal ATP by administering apyrase produces a reduction in withdrawal behaviors. Conversely, we found that pharmacological inhibition of ATP breakdown exacerbates withdrawal. Treatment with a Panx1-blocking peptide (10panx) or the clinically used broad-spectrum Panx1 blockers, mefloquine or probenecid, suppressed ATP release and reduced withdrawal severity. Our results demonstrate that Panx1-mediated ATP release from microglia is required for morphine withdrawal in rodents and that blocking Panx1 alleviates the severity of withdrawal without affecting opiate analgesia.

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Figure 1: Repeated morphine treatment increases microglial Panx1 expression and activity.
Figure 2: Blocking microglial Panx1 alleviates morphine withdrawal.
Figure 3: Genetic ablation of microglial Panx1 prevents naloxone-induced synaptic facilitation in spinal lamina I–II dorsal horn neurons of morphine-treated mice.
Figure 4: ATP released from microglial Panx1 is suppressed by mefloquine and probenecid.

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  • 15 February 2017

    In the version of this article initially published online, Yves De Koninck’s name was misspelled in the author list. The original version listed Yves DeKoninck. The error has been corrected in the print, PDF and HTML versions of this article.

References

  1. Stotts, A.L. et al. A stage I pilot study of acceptance and commitment therapy for methadone detoxification. Drug Alcohol Depend. 125, 215–222 (2012).

    Article  Google Scholar 

  2. Weiss, R.D. et al. Reasons for opioid use among patients with dependence on prescription opioids: the role of chronic pain. J. Subst. Abuse Treat. 47, 140–145 (2014).

    Article  Google Scholar 

  3. Frank, J.W. et al. Patients' perspectives on tapering of chronic opioid therapy: a qualitative study. Pain Med. 17, 1838–1847 (2016).

    Article  Google Scholar 

  4. Yarborough, B.J.H. et al. Methadone, buprenorphine and preferences for opioid agonist treatment: a qualitative analysis. Drug Alcohol Depend. 160, 112–118 (2016).

    Article  CAS  Google Scholar 

  5. Pelegrin, P. & Surprenant, A. Pannexin-1 mediates large pore formation and interleukin-1β release by the ATP-gated P2X7 receptor. EMBO J. 25, 5071–5082 (2006).

    Article  CAS  Google Scholar 

  6. Sandilos, J.K. et al. Pannexin 1, an ATP release channel, is activated by caspase cleavage of its pore-associated C-terminal autoinhibitory region. J. Biol. Chem. 287, 11303–11311 (2012).

    Article  CAS  Google Scholar 

  7. Huang, Y., Grinspan, J.B., Abrams, C.K. & Scherer, S.S. Pannexin1 is expressed by neurons and glia but does not form functional gap junctions. Glia 55, 46–56 (2007).

    Article  Google Scholar 

  8. Iglesias, R., Dahl, G., Qiu, F., Spray, D.C. & Scemes, E. Pannexin 1: the molecular substrate of astrocyte “hemichannels”. J. Neurosci. 29, 7092–7097 (2009).

    Article  CAS  Google Scholar 

  9. Iglesias, R. et al. P2X7 receptor–pannexin1 complex: pharmacology and signaling. Am. J. Physiol. Cell Physiol. 295, C752–C760 (2008).

    Article  CAS  Google Scholar 

  10. Sorge, R.E. et al. Genetically determined P2X7 receptor pore formation regulates variability in chronic pain sensitivity. Nat. Med. 18, 595–599 (2012).

    Article  CAS  Google Scholar 

  11. 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 

  12. Masuda, T. et al. Transcription factor IRF5 drives P2X4R+-reactive microglia gating neuropathic pain. Nat. Commun. 5, 3771 (2014).

    Article  CAS  Google Scholar 

  13. Parkhurst, C.N. et al. Microglia promote learning-dependent synapse formation through brain-derived neurotrophic factor. Cell 155, 1596–1609 (2013).

    Article  CAS  Google Scholar 

  14. Bagley, E.E. et al. Drug-induced GABA transporter currents enhance GABA release to induce opioid withdrawal behaviors. Nat. Neurosci. 14, 1548–1554 (2011).

    Article  CAS  Google Scholar 

  15. Bonin, R.P. & De Koninck, Y. A spinal analog of memory reconsolidation enables reversal of hyperalgesia. Nat. Neurosci. 17, 1043–1045 (2014).

    Article  CAS  Google Scholar 

  16. Huang, Y.-J. et al. The role of pannexin 1 hemichannels in ATP release and cell–cell communication in mouse taste buds. Proc. Natl. Acad. Sci. USA 104, 6436–6441 (2007).

    Article  CAS  Google Scholar 

  17. Chiang, C.Y. et al. Endogenous ATP involvement in mustard-oil-induced central sensitization in trigeminal subnucleus caudalis (medullary dorsal horn). J. Neurophysiol. 94, 1751–1760 (2005).

    Article  CAS  Google Scholar 

  18. Beckel, J.M. et al. Pannexin 1 channels mediate the release of ATP into the lumen of the rat urinary bladder. J. Physiol. (Lond.) 593, 1857–1871 (2015).

    Article  CAS  Google Scholar 

  19. Nakatsuka, T. & Gu, J.G. ATP P2X receptor–mediated enhancement of glutamate release and evoked EPSCs in dorsal horn neurons of the rat spinal cord. J. Neurosci. 21, 6522–6531 (2001).

    Article  CAS  Google Scholar 

  20. 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 

  21. Iglesias, R., Spray, D.C. & Scemes, E. Mefloquine blockade of pannexin1 currents: resolution of a conflict. Cell Commun. Adhes. 16, 131–137 (2009).

    Article  CAS  Google Scholar 

  22. Drdla, R., Gassner, M., Gingl, E. & Sandkühler, J. Induction of synaptic long-term potentiation after opioid withdrawal. Science 325, 207–210 (2009).

    Article  CAS  Google Scholar 

  23. Han, M.-H. et al. Role of cAMP response element–binding protein in the rat locus ceruleus: regulation of neuronal activity and opiate withdrawal behaviors. J. Neurosci. 26, 4624–4629 (2006).

    Article  CAS  Google Scholar 

  24. Koo, J.W. et al. Epigenetic basis of opiate suppression of Bdnf gene expression in the ventral tegmental area. Nat. Neurosci. 18, 415–422 (2015).

    Article  CAS  Google Scholar 

  25. Vargas-Perez, H. et al. Ventral tegmental area BDNF induces an opiate-dependent-like reward state in naive rats. Science 324, 1732–1734 (2009).

    Article  CAS  Google Scholar 

  26. Laviolette, S.R., Gallegos, R.A., Henriksen, S.J. & van der Kooy, D. Opiate state controls bi-directional reward signaling via GABAA receptors in the ventral tegmental area. Nat. Neurosci. 7, 160–169 (2004).

    Article  CAS  Google Scholar 

  27. Zhu, Y., Wienecke, C.F.R., Nachtrab, G. & Chen, X. A thalamic input to the nucleus accumbens mediates opiate dependence. Nature 530, 219–222 (2016).

    Article  CAS  Google Scholar 

  28. Trang, T., Ma, W., Chabot, J.-G., Quirion, R. & Jhamandas, K. Spinal modulation of calcitonin gene–related peptide by endocannabinoids in the development of opioid physical dependence. Pain 126, 256–271 (2006).

    Article  CAS  Google Scholar 

  29. Ferrini, F. et al. Morphine hyperalgesia gated through microglia-mediated disruption of neuronal Cl homeostasis. Nat. Neurosci. 16, 183–192 (2013).

    Article  CAS  Google Scholar 

  30. Ichikizaki, K., Toya, S. & Hoshino, T. A new procedure for lumbar puncture in the mouse (intrathecal injection) preliminary report. Keio J. Med. 28, 165–171 (1979).

    Article  CAS  Google Scholar 

  31. Oyebamiji, A.I. et al. Characterization of migration parameters on peripheral and central nervous system T cells following treatment of experimental allergic encephalomyelitis with CRYAB. J. Neuroimmunol. 259, 66–74 (2013).

    Article  CAS  Google Scholar 

  32. Trang, T., Beggs, S., Wan, X. & Salter, M.W. P2X4-receptor-mediated synthesis and release of brain-derived neurotrophic factor in microglia is dependent on calcium and p38-mitogen-activated protein kinase activation. J. Neurosci. 29, 3518–3528 (2009).

    Article  CAS  Google Scholar 

  33. Weilinger, N.L., Tang, P.L. & Thompson, R.J. Anoxia-induced NMDA receptor activation opens pannexin channels via Src family kinases. J. Neurosci. 32, 12579–12588 (2012).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank D. Littman and W.-B. Gan (both at New York University School of Medicine) for generously providing breeding pairs for the Cx3cr1-CreERT2 mouse colony, R. Thompson (University of Calgary) for providing the Panx1flx/flx mice, and F. Visser for mouse genotyping. BV-2 microglial-like cells were provided by M. Tsuda (Kyushu University) and K. Biber (University of Groningen). We also thank K. Jhamandas and B. Zochodne for comments on the manuscript and the RUN CORE Facility for access to the Nikon C1S1 confocal and A1R multiphoton microscopes. This work was supported by grants from the Vi Riddell Program for Pediatric Pain, Natural Sciences and Engineering Research Council of Canada (RGPIN418299) and the Rita Allen Foundation and American Pain Society to T.T. Canadian Institutes of Health Research grants were also awarded to T.T. (MOP133523), Y.D.K. (MOP12942) and G.W.Z. (FDN143336). N.E.B. is supported by a Hotchkiss Brain Institute Doctoral Scholarship and a Queen Elizabeth II Scholarship. R.P.B., G.W.Z. and Y.D.K. hold Canada Research Chairs.

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N.E.B. and T.T. conceived and designed the project. N.E.B., R.P.B., H.L.-P., C.B., Z.F.C., M.M., J.V.S., P.L.S., D.B. and C.M.C. performed the experiments. T.T., Y.D.K., S.L.B., M.C.A., G.W.Z. and J.S.B. supervised the experiments. N.E.B., R.P.B., H.L.-P., M.C.A. and C.M.C. analyzed the data. N.E.B. and T.T. wrote the manuscript. All authors read and approved the manuscript.

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Correspondence to Tuan Trang.

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Burma, N., Bonin, R., Leduc-Pessah, H. et al. Blocking microglial pannexin-1 channels alleviates morphine withdrawal in rodents. Nat Med 23, 355–360 (2017). https://doi.org/10.1038/nm.4281

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