Assembly and architecture of endogenous NMDA receptors in adult cerebral cortex and hippocampus

Ming Zhang; Juan Feng; Chun Xie; Nan Song; Chaozhi Jin; Jian Wang; Qun Zhao; Lihua Zhang; Boshuang Wang; Yidi Sun; Fei Guo; Yang Li; Shujia Zhu

doi:10.1016/j.cell.2025.01.004

Assembly and architecture of endogenous NMDA receptors in adult cerebral cortex and hippocampus

Cell. 2025 Jan 17:S0092-8674(25)00033-9. doi: 10.1016/j.cell.2025.01.004. Online ahead of print.

Authors

Ming Zhang¹, Juan Feng², Chun Xie², Nan Song², Chaozhi Jin³, Jian Wang³, Qun Zhao⁴, Lihua Zhang⁴, Boshuang Wang⁵, Yidi Sun², Fei Guo¹, Yang Li⁶, Shujia Zhu⁷

Affiliations

¹ Center for Neurological and Psychiatric Research and Drug Discovery, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; University of Chinese Academy of Sciences, Beijing, China.
² Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
³ State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 102206, China.
⁴ CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, State Key Laboratory of Medical Proteomics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
⁵ University of Chinese Academy of Sciences, Beijing, China; Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
⁶ Center for Neurological and Psychiatric Research and Drug Discovery, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; University of Chinese Academy of Sciences, Beijing, China. Electronic address: liyang@simm.ac.cn.
⁷ University of Chinese Academy of Sciences, Beijing, China; Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: shujiazhu@ion.ac.cn.

PMID: 39855198
DOI: 10.1016/j.cell.2025.01.004

Abstract

The cerebral cortex and hippocampus are crucial brain regions for learning and memory, which depend on activity-induced synaptic plasticity involving N-methyl-ᴅ-aspartate receptors (NMDARs). However, subunit assembly and molecular architecture of endogenous NMDARs (eNMDARs) in the brain remain elusive. Using conformation- and subunit-dependent antibodies, we purified eNMDARs from adult rat cerebral cortex and hippocampus. Three major subtypes of GluN1-N2A-N2B, GluN1-N2B, and GluN1-N2A eNMDARs were resolved by cryoelectron microscopy (cryo-EM) at the resolution up to 4.2 Å. The particle ratio of these three subtypes was 9:7:4, indicating that about half of GluN2A and GluN2B subunits are incorporated into the tri-heterotetramers. Structural analysis revealed the asymmetric architecture of the GluN1-N2A-N2B receptor throughout the extracellular to the transmembrane layers. Moreover, the conformational variations between GluN1-N2B and GluN1-N2A-N2B receptors revealed the distinct biophysical properties across different eNMDAR subtypes. Our findings imply the structural and functional complexity of eNMDARs and shed light on structure-based therapeutic design targeting these eNMDARs in vivo.

Keywords: GluN1-N2A receptor; GluN1-N2A-N2B receptor; GluN1-N2B receptor; cortex and hippocampus; cryo-EM; endogenous NMDA receptor; learning and memory; receptor pharmacology.