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Morphine hyperalgesia gated through microglia-mediated disruption of neuronal Cl− homeostasis

Nature Neuroscience volume 16, pages 183–192 (2013)Cite this article

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Abstract

A major unresolved issue in treating pain is the paradoxical hyperalgesia produced by the gold-standard analgesic morphine and other opiates. We found that hyperalgesia-inducing treatment with morphine resulted in downregulation of the K+-Cl− co-transporter KCC2, impairing Cl− homeostasis in rat spinal lamina l neurons. Restoring the anion equilibrium potential reversed the morphine-induced hyperalgesia without affecting tolerance. The hyperalgesia was also reversed by ablating spinal microglia. Morphine hyperalgesia, but not tolerance, required μ opioid receptor–dependent expression of P2X4 receptors (P2X4Rs) in microglia and μ-independent gating of the release of brain-derived neurotrophic factor (BDNF) by P2X4Rs. Blocking BDNF-TrkB signaling preserved Cl− homeostasis and reversed the hyperalgesia. Gene-targeted mice in which Bdnf was deleted from microglia did not develop hyperalgesia to morphine. However, neither morphine antinociception nor tolerance was affected in these mice. Our findings dissociate morphine-induced hyperalgesia from tolerance and suggest the microglia-to-neuron P2X4-BDNF-KCC2 pathway as a therapeutic target for preventing hyperalgesia without affecting morphine analgesia.

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Figure 1: Repeated morphine administration causes hyperalgesia and tolerance.
Figure 2: Morphine disrupts Cl− homeostasis in lamina I neurons.
Figure 3: Morphine effects on activity and expression of KCC2.
Figure 4: Morphine-induced hyperalgesia depends on microglia activation.
Figure 5: P2X4Rs in microglia are required for morphine-induced hyperalgesia.
Figure 6: Altered Cl− homeostasis in spinal neurons and morphine-induced hyperalgesia are dependent on P2X4R-BDNF-TrkB signaling.
Figure 7: Genetic deletion of Bdnf from microglia abrogates development of morphine-induced hyperalgesia, but not tolerance.
Figure 8: Activation of two separate signaling pathways is necessary for morphine-induced hyperalgesia.

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Acknowledgements

We thank K. Bachand for technical assistance with behavior, F. Rassendren (INSERM) for P2rx4−/− mice, C.J. Evans (National Institute on Drug Abuse) for (+)naloxone and J. Vlahakis for (+)naloxone polarimetric analysis. This paper is dedicated to the memory of Karen Vandal, who passed away during the course of this study. This work was supported by Canadian Institutes for Health Research (CIHR) grants to Y.D.K., M.W.S., C.M.C. and J.-M.B., by the Krembil Foundation (Y.D.K. and M.W.S.), the Regione Piemonte/University of Turin Fellowship Program (F.F.), the CIHR Fellowship program (T.T. and L.-E.L.), the Howard Hughes Medical Institute (M.W.S.), the Anne and Max Tanenbaum Chair Program (M.W.S.), the Canada Research Chairs Program (C.M.C., J.-M.B. and M.W.S.), the Fonds de la recherche en santé du Québec Chercheur National Program (Y.D.K.) and the Ontario Research Fund Research Excellence Program (M.W.S.).

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  1. Francesco Ferrini and Tuan Trang: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut Universitaire en Santé Mentale de Québec, Québec, Canada

    Francesco Ferrini, Sophie Laffray, Thomas Del'Guidice, Louis-Etienne Lorenzo, Annie Castonguay, Nicolas Doyon, Antoine G Godin, Karen Vandal, Jean-Martin Beaulieu & Yves De Koninck

  2. Department of Psychiatry and Neuroscience, Université Laval, Québec, Québec, Canada

    Francesco Ferrini, Sophie Laffray, Thomas Del'Guidice, Louis-Etienne Lorenzo, Annie Castonguay, Nicolas Doyon, Antoine G Godin, Jean-Martin Beaulieu & Yves De Koninck

  3. Department of Veterinary Sciences, University of Turin, Turin, Italy

    Francesco Ferrini

  4. Program in Neurosciences & Mental Health, Hospital for Sick Children, Toronto, Ontario, Canada

    Tuan Trang, Wenbo Zhang, Daniela Mohr, Simon Beggs & Michael W Salter

  5. Department of Physiology, University of Toronto, Toronto, Ontario, Canada

    Tuan Trang, Wenbo Zhang, Daniela Mohr, Simon Beggs & Michael W Salter

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

    Tuan Trang

  7. Department of Physiology and Pharmacology, Hotchkiss Brain Institute, University of Calgary, Calgary, Canada

    Tuan Trang

  8. Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada

    Theresa-Alexandra M Mattioli & Catherine M Cahill

  9. Department of Anesthesiology and Perioperative Care, University of California, Irvine, California, USA

    Catherine M Cahill

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  1. Francesco Ferrini
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Contributions

F.F., T.T., C.M.C., M.W.S. and Y.D.K. conceived and designed the project. C.M.C., J.-M.B., Y.D.K. and M.W.S. supervised the experiments. F.F., T.T., T.-A.M.M., S.L., T.D., L.-E.L., A.C., W.Z., D.M., S.B. and K.V. performed the experiments. N.D. performed computer simulations and contributed to interpretation of results. S.B. generated CD11b-cre; BdnfloxP/loxP mice. F.F., T.T., T.-A.M.M., S.L., T.D., L.-E.L., A.C., N.D., W.Z. and A.G.G. analyzed the data. F.F., T.T., M.W.S. and Y.D.K. wrote the manuscript. All of the authors read and discussed the manuscript.

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Correspondence to Yves De Koninck.

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Ferrini, F., Trang, T., Mattioli, TA. et al. Morphine hyperalgesia gated through microglia-mediated disruption of neuronal Cl− homeostasis. Nat Neurosci 16, 183–192 (2013). https://doi.org/10.1038/nn.3295

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