• An Erratum to this article was published on 06 June 2017

This article has been updated


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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Change history

  • 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.


  1. 1.

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

  2. 2.

    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).

  3. 3.

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

  4. 4.

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

  5. 5.

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

  6. 6.

    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).

  7. 7.

    , , & Pannexin1 is expressed by neurons and glia but does not form functional gap junctions. Glia 55, 46–56 (2007).

  8. 8.

    , , , & Pannexin 1: the molecular substrate of astrocyte “hemichannels”. J. Neurosci. 29, 7092–7097 (2009).

  9. 9.

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

  10. 10.

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

  11. 11.

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

  12. 12.

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

  13. 13.

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

  14. 14.

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

  15. 15.

    & A spinal analog of memory reconsolidation enables reversal of hyperalgesia. Nat. Neurosci. 17, 1043–1045 (2014).

  16. 16.

    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).

  17. 17.

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

  18. 18.

    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).

  19. 19.

    & 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).

  20. 20.

    , & Probenecid, a gout remedy, inhibits pannexin 1 channels. Am. J. Physiol. Cell Physiol. 295, C761–C767 (2008).

  21. 21.

    , & Mefloquine blockade of pannexin1 currents: resolution of a conflict. Cell Commun. Adhes. 16, 131–137 (2009).

  22. 22.

    , , & Induction of synaptic long-term potentiation after opioid withdrawal. Science 325, 207–210 (2009).

  23. 23.

    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).

  24. 24.

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

  25. 25.

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

  26. 26.

    , , & Opiate state controls bi-directional reward signaling via GABAA receptors in the ventral tegmental area. Nat. Neurosci. 7, 160–169 (2004).

  27. 27.

    , , & A thalamic input to the nucleus accumbens mediates opiate dependence. Nature 530, 219–222 (2016).

  28. 28.

    , , , & Spinal modulation of calcitonin gene–related peptide by endocannabinoids in the development of opioid physical dependence. Pain 126, 256–271 (2006).

  29. 29.

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

  30. 30.

    , & A new procedure for lumbar puncture in the mouse (intrathecal injection) preliminary report. Keio J. Med. 28, 165–171 (1979).

  31. 31.

    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).

  32. 32.

    , , & 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).

  33. 33.

    , & Anoxia-induced NMDA receptor activation opens pannexin channels via Src family kinases. J. Neurosci. 32, 12579–12588 (2012).

Download references


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.

Author information


  1. Department of Comparative Biology and Experimental Medicine, University of Calgary, Calgary, Alberta, Canada.

    • Nicole E Burma
    • , Heather Leduc-Pessah
    • , Zoe F Cairncross
    • , Michael Mousseau
    •  & Tuan Trang
  2. Department of Physiology and Pharmacology, Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada.

    • Nicole E Burma
    • , Heather Leduc-Pessah
    • , Corey Baimel
    • , Zoe F Cairncross
    • , Michael Mousseau
    • , Patrick L Stemkowski
    • , Dinara Baimoukhametova
    • , Jaideep S Bains
    • , Michael C Antle
    • , Gerald W Zamponi
    • , Stephanie L Borgland
    •  & Tuan Trang
  3. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada.

    • Robert P Bonin
  4. Department of Psychology, University of Calgary, Calgary, Alberta, Canada.

    • Jhenkruthi Vijaya Shankara
    •  & Michael C Antle
  5. Department of Anesthesiology and Perioperative Care, University of California Irvine, Irvine, California, USA.

    • Catherine M Cahill
  6. Department of Psychiatry and Neuroscience, Institut Universitaire en santé mentale de Québec, Université Laval, Ville de Québec, Québec, Canada.

    • Yves De Koninck


  1. Search for Nicole E Burma in:

  2. Search for Robert P Bonin in:

  3. Search for Heather Leduc-Pessah in:

  4. Search for Corey Baimel in:

  5. Search for Zoe F Cairncross in:

  6. Search for Michael Mousseau in:

  7. Search for Jhenkruthi Vijaya Shankara in:

  8. Search for Patrick L Stemkowski in:

  9. Search for Dinara Baimoukhametova in:

  10. Search for Jaideep S Bains in:

  11. Search for Michael C Antle in:

  12. Search for Gerald W Zamponi in:

  13. Search for Catherine M Cahill in:

  14. Search for Stephanie L Borgland in:

  15. Search for Yves De Koninck in:

  16. Search for Tuan Trang in:


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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Tuan Trang.

Supplementary information

PDF files

  1. 1.

    Supplementary Figures

    Supplementary Figures 1–10

About this article

Publication history






Further reading