Abstract
Ethanol actions on α-amino-3-hydyroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors were studied using voltage-clamp recordings from mouse cortical and hippocampal neurons. During whole-cell recordings ethanol (EtOH) inhibited AMPA receptor-mediated currents in a dose-dependent manner at concentrations from 10 to 500 mM. The steady-state component of AMPA-activated current was more sensitive to EtOH than the peak component. To examine the effect of EtOH on a well resolved peak current component, patches were excised from cultured cortical neurons, to which AMPA and EtOH were applied using a piezoelectric solution application system. Under this condition, the peak current was not inhibited significantly by EtOH. To further study possible mechanisms of EtOH inhibition, kainate and AMPA were used to evoke currents in the absence and presence of cyclothiazide. Ethanol inhibition was stronger when receptors were activated by low than high kainate concentrations. Cyclothiazide reduced inhibition by EtOH regardless of the agonist used to activate the receptor. Finally, EtOH inhibition was reduced in a point mutated (L497Y) GluRAi receptor that lacks desensitization. These findings suggest that EtOH inhibits AMPA receptors by stabilizing the desensitized state. Our results can explain some of the variation observed in EtOH inhibition in previous studies, and support the idea that physiologically relevant concentrations of EtOH can have a strong effect on AMPA receptor function.
Footnotes
-
This study was supported by AA08986 (to D.M.L. and T.M.), the Sigrid Juselius Foundation (to D.M.L. and E.R.K.), the Finnish Foundation for Alcohol Studies (to T.M. and E.R.K.), and the National Institute on Alcohol Abuse and Alcoholism Division of Intramural Clinical and Basic Research.
-
DOI: 10.1124/jpet.103.050666.
-
ABBREVIATIONS: CNS, central nervous system; NMDA, N-methyl-d-aspartate; AMPA, α-amino-3-hydyroxy-5-methyl-4-isoxazolepropionic acid; AMPAR, AMPA receptor; EtOH, ethanol; HEK, human embryonic kidney; EGFP, enhanced green fluorescent protein; ANOVA, analysis of variance; Con A, concanavalin A; WT, wild-type.
- Received February 19, 2003.
- Accepted May 6, 2003.
- The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|