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Glycine binding primes NMDA receptor internalization

Abstract

NMDA (N-methyl-d-aspartate) receptors (NMDARs) are a principal subtype of excitatory ligand-gated ion channel with prominent roles in physiological and disease processes in the central nervous system1. Recognition that glycine potentiates NMDAR-mediated currents2 as well as being a requisite co-agonist of the NMDAR subtype of ‘glutamate’ receptor3 profoundly changed our understanding of chemical synaptic communication in the central nervous system. The binding of both glycine and glutamate is necessary to cause opening of the NMDAR conductance pore1. Although binding of either agonist alone is insufficient to cause current flow through the channel, we report here that stimulation of the glycine site initiates signalling through the NMDAR complex, priming the receptors for clathrin-dependent endocytosis. Glycine binding alone does not cause the receptor to be endocytosed; this requires both glycine and glutamate site activation of NMDARs. The priming effect of glycine is mimicked by the NMDAR glycine site agonist d-serine, and is blocked by competitive glycine site antagonists. Synaptic as well as extrasynaptic NMDARs are primed for internalization by glycine site stimulation. Our results demonstrate transmembrane signal transduction through activating the glycine site of NMDARs, and elucidate a model for modulating cell–cell communication in the central nervous system.

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Figure 1: Co-agonist stimulation induces dynamin-dependent NMDA receptor internalization.
Figure 2: Glycine site stimulation initiates a progressive decline in NMDA receptor-mediated currents.
Figure 3: Glycine binding primes the decline of NMDA responses and recruits adaptin β2 to the NMDA receptor complex.
Figure 4: Glycine primes dynamin-dependent internalization of NMDA receptors and leads to preferential suppression of NMDAR- but not AMPAR-mediated synaptic currents.

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Acknowledgements

We thank J. L. Hicks and Y. Li for technical support, and J. F. MacDonald and L. Y. Wang for comments on the manuscript. This work was supported by the Canadian Institutes of Health Research (CIHR; to M.W.S. and Y.T.W.), the EJLB Foundation (to Y.T.W.), the Ontario Neurotrauma Foundation (ONF; to M.W.S.) and the Nicole Fealdman Memorial Fund (to M.W.S.). M.W.S. is a CIHR Investigator, Y.T.W. is a CIHR Investigator and the holder of the HSF of British Columbia and Yukon Chair in Stroke Research at the University of British Columbia and Vancouver Hospital and Health Sciences Centre, and is an International Scholar of the Howard Hughes Medical Institute. Y.N. is a Ronald Melzack Fellow of CIHR; Y.Q.H. is a Fellow of the ONF and CIHR/Heart and Stroke Foundation of Canada (HSFC); G.A. is a Fellow of Research Training Centre at Hospital for Sick Children; W.J. is a CIHR/HSFC student; and L.V.K. is a CIHR MD/PhD student. We thank J. F. MacDonald and M. Sheng for providing cDNA constructs. Y.T.W. and M.W.S. are joint senior authors.

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Correspondence to Yu Tian Wang or Michael W. Salter.

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Nong, Y., Huang, YQ., Ju, W. et al. Glycine binding primes NMDA receptor internalization. Nature 422, 302–307 (2003). https://doi.org/10.1038/nature01497

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