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Excitatory glycine receptors containing the NR3 family of NMDA receptor subunits

Abstract

The N-methyl-d-aspartate subtype of glutamate receptor (NMDAR) serves critical functions in physiological and pathological processes in the central nervous system, including neuronal development, plasticity and neurodegeneration1,2. Conventional heteromeric NMDARs composed of NR1 and NR2A–D subunits3,4 require dual agonists, glutamate and glycine, for activation. They are also highly permeable to Ca2+, and exhibit voltage-dependent inhibition by Mg2+. Coexpression of NR3A with NR1 and NR2 subunits modulates NMDAR activity5,6,7. Here we report the cloning and characterization of the final member of the NMDAR family, NR3B, which shares high sequence homology with NR3A. From in situ and immunocytochemical analyses, NR3B is expressed predominantly in motor neurons, whereas NR3A is more widely distributed5,6. Remarkably, when co-expressed in Xenopus oocytes, NR3A or NR3B co-assembles with NR1 to form excitatory glycine receptors that are unaffected by glutamate or NMDA, and inhibited by d-serine, a co-activator of conventional NMDARs. Moreover, NR1/NR3A or -3B receptors form relatively Ca2+-impermeable cation channels that are resistant to Mg2+, MK-801, memantine and competitive antagonists. In cerebrocortical neurons containing NR3 family members, glycine triggers a burst of firing, and membrane patches manifest glycine-responsive single channels that are suppressible by d-serine. By itself, glycine is normally thought of as an inhibitory neurotransmitter. In contrast, these NR1/NR3A or -3B ‘NMDARs’ constitute a type of excitatory glycine receptor.

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Figure 1: NR3B expression pattern by in situ hybridization and immunocytochemistry.
Figure 2: Pharmacological characterization of NR1/NR3B or -3A receptors in Xenopus oocytes.
Figure 3: Ion selectivity (ad) and single-channel analysis (eg) of NR1/NR3B receptors in Xenopus oocytes.
Figure 4: Physiological responses to glycine in rat cerebrocortical neurons.

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Acknowledgements

We thank H.-S. V. Chen for discussions and T. Ciabarra and M. Forcina for preliminary work. Monoclonal antibody K35/40 was made in collaboration with J. S. Trimmer and N. J. Sucher. NR1 and NR2A cDNAs were gifts of S. F. Heinemann and S. Nakanishi, respectively. This work was supported in part by grants from the National Institutes of Health. S.A.L. and D.Z. are co-senior authors.

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Correspondence to Stuart A. Lipton or Dongxian Zhang.

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The authors have a patent application pending regarding NR3 subunits.

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Chatterton, J., Awobuluyi, M., Premkumar, L. et al. Excitatory glycine receptors containing the NR3 family of NMDA receptor subunits. Nature 415, 793–798 (2002). https://doi.org/10.1038/nature715

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