Specific pathogenic mutations in the M3 domain of the GluN1 subunit regulate the surface delivery and pharmacological sensitivity of NMDA receptors
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
33773999
DOI
10.1016/j.neuropharm.2021.108528
PII: S0028-3908(21)00082-4
Knihovny.cz E-resources
- Keywords
- Electrophysiology, Glutamate receptor, Hippocampal neuron, Ion channel, Memantine, Membrane domain,
- MeSH
- Excitatory Amino Acid Agonists administration & dosage MeSH
- Excitatory Amino Acid Antagonists administration & dosage MeSH
- HEK293 Cells MeSH
- Hippocampus drug effects physiology MeSH
- Rats MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Mutation drug effects genetics MeSH
- Protein Subunits agonists antagonists & inhibitors genetics MeSH
- Rats, Wistar MeSH
- Surface Properties drug effects MeSH
- Nerve Tissue Proteins agonists antagonists & inhibitors genetics MeSH
- Receptors, N-Methyl-D-Aspartate agonists antagonists & inhibitors genetics MeSH
- Dose-Response Relationship, Drug MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Excitatory Amino Acid Agonists MeSH
- Excitatory Amino Acid Antagonists MeSH
- GRIN1 protein, human MeSH Browser
- Protein Subunits MeSH
- Nerve Tissue Proteins MeSH
- Receptors, N-Methyl-D-Aspartate MeSH
N-methyl-d-aspartate receptors (NMDARs) play an essential role in regulating glutamatergic neurotransmission. Recently, pathogenic missense mutations were identified in genes encoding NMDAR subunits; however, their effect on NMDAR activity is often poorly understood. Here, we examined whether three previously identified pathogenic mutations (M641I, A645S, and Y647S) in the M3 domain of the GluN1 subunit affect the receptor's surface delivery, agonist sensitivity, Mg2+ block, and/or inhibition by the FDA-approved NMDAR blocker memantine. When expressed in HEK293 cells, we found reduced surface expression of GluN1-M641I/GluN2A, GluN1-Y647S/GluN2A, and GluN1-Y647S/GluN2B receptors; other mutation-bearing NMDAR combinations, including GluN1/GluN3A receptors, were expressed at normal surface levels. When expressed in rat hippocampal neurons, we consistently found reduced surface expression of the GluN1-M641I and GluN1-Y647S subunits when compared with wild-type GluN1 subunit. At the functional level, we found that GluN1-M641I/GluN2 and GluN1-A645S/GluN2 receptors expressed in HEK293 cells have wild-type EC50 values for both glutamate and glycine; in contrast, GluN1-Y647S/GluN2 receptors do not produce glutamate-induced currents. In the presence of a physiological concentration of Mg2+, we found that GluN1-M641I/GluN2 receptors have a lower memantine IC50 and slower offset kinetics, whereas GluN1-A645S/GluN2 receptors have a higher memantine IC50 and faster offset kinetics when compared to wild-type receptors. Finally, we found that memantine was the most neuroprotective in hippocampal neurons expressing GluN1-M641I subunits, followed by neurons expressing wild-type GluN1 and then GluN1-A645S subunits in an NMDA-induced excitotoxicity assay. These results indicate that specific pathogenic mutations in the M3 domain of the GluN1 subunit differentially affect the trafficking and functional properties of NMDARs.
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