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Zinc alleviates pain through high-affinity binding to the NMDA receptor NR2A subunit

Abstract

Zinc is abundant in the central nervous system and regulates pain, but the underlying mechanisms are unknown. In vitro studies have shown that extracellular zinc modulates a plethora of signaling membrane proteins, including NMDA receptors containing the NR2A subunit, which display exquisite zinc sensitivity. We created NR2A-H128S knock-in mice to investigate whether Zn2+–NR2A interaction influences pain control. In these mice, high-affinity (nanomolar) zinc inhibition of NMDA currents was lost in the hippocampus and spinal cord. Knock-in mice showed hypersensitivity to radiant heat and capsaicin, and developed enhanced allodynia in inflammatory and neuropathic pain models. Furthermore, zinc-induced analgesia was completely abolished under both acute and chronic pain conditions. Our data establish that zinc is an endogenous modulator of excitatory neurotransmission in vivo and identify a new mechanism in pain processing that relies on NR2A NMDA receptors. The study also potentially provides a molecular basis for the pain-relieving effects of dietary zinc supplementation.

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Figure 1: Targeting the NMDA receptor NR2A subunit gene in mice.
Figure 2: High-affinity zinc inhibition of NMDA currents is lost in NR2A-H128S mice.
Figure 3: NR2A-H128S mice show enhanced basal pain sensitivity in response to radiant heat and capsaicin.
Figure 4: NR2A-H128S mice show increased mechanical allodynia under chronic pain.
Figure 5: Zinc analgesia is abolished in NR2A-H128S mice.

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Acknowledgements

We thank B. Puvion, A. Matifas and R. Weibel for technical assistance and E. Krejci for help with the targeting vector. We thank B. Barbour, J. Becker and E. Schwartz for comments on the manuscript. We also thank Victor Faundez (Emory University) for the gift of the ZnT3 antibody. This research was supported by the Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM), the Université de Strasbourg, the Agence Nationale de la Recherche (France) grant SynapticZinc (P.P. and B.K.), the US National Institutes of Health, National Institue on Drug Abuse grant DA05010 (B.K.) and an Equipe Fondation pour la Recherche Médicale (FRM) grant (P.P.). C.N. was supported by the FRM and A.M.V. by the Région Ile-de-France.

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B.L.K. and P.P. designed and supervised the study. D.F., J.N. and A.L.G. contributed to the creation NR2A-H128S mutant mouse line. J.N. and P.P. performed the electrophysiologial experiments on Xenopus oocytes. A.M.V. ran electrophysiological characterization of mutant mice and performed the Timm's staining. C.N. performed all of the pain experiments. D.R. and A.-M.O. conducted neurological examination of mutant mice. S.C. and A.M.V. performed immunochemistry. C.G.-R. and J.N. contributed to conceptual aspects of the study. C.N., A.M.V., C.G.-R., A.-M.O., P.P. and B.L.K. wrote the manuscript.

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Correspondence to Pierre Paoletti.

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Nozaki, C., Vergnano, A., Filliol, D. et al. Zinc alleviates pain through high-affinity binding to the NMDA receptor NR2A subunit. Nat Neurosci 14, 1017–1022 (2011). https://doi.org/10.1038/nn.2844

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