Reduced Ethanol Inhibition ofN-Methyl-d-aspartate Receptors by Deletion of the NR1 C0 Domain or Overexpression of α-Actinin-2 Proteins

• Published in: The Journal of biological chemistry Vol. 275; no. 20; pp. 15019 - 15024
• Main Authors: Anders, Douglas L., Blevins, Tana, Smothers, Corigan T., Woodward, John J.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 19.05.2000
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.275.20.15019

The depressant actions of ethanol on central nervous system activity appear to be mediated by its actions on a number of important membrane associated ion channels including theN-methyl-d-aspartate (NMDA) subtype of ionotropic glutamate receptor. Although no specific site of action for ethanol on the NMDA receptor has been found, previous studies suggest that the ethanol sensitivity of the receptor may be affected by intracellular C-terminal domains of the receptor that regulate the calcium-dependent inactivation of the receptor. In the present study, co-expression of the NR2A subunit and an NR1 subunit that lacks the alternatively spliced intracellular C1 cassette did not reduce the effects of ethanol on channel function as measured by patch-clamp electrophysiology. Full inhibition was also observed in cells expressing an NR1 subunit truncated at the end of the C0 domain (NR1863stop). However, the inhibitory effects of ethanol were reduced by expression of an NR1 C0 domain deletion mutant (NR1Δ839–863), truncation mutant (NR1858stop), or a triple-point mutant (Arg to Ala, Lys to Ala, and Asn to Ala at 859–861) previously shown to significantly reduce calcium-dependent inactivation. A similar reduction in the effects of ethanol on wild-type NR1/2A but not NR1/2B or NR1/2C receptors was observed after co-expression of full-length or truncated human skeletal muscle α-actinin-2 proteins that produce a functional knockout of the C0 domain. The effects of ethanol on hippocampal and cortical NMDA-induced currents were similarly attenuated in low calcium recording conditions, suggesting that a C0 domain-dependent process may confer additional ethanol sensitivity to NMDA receptors.