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Originally published In Press as doi:10.1074/jbc.M100317200 on May 31, 2001

J. Biol. Chem., Vol. 276, Issue 32, 29764-29771, August 10, 2001
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Differential Effects of Chronic Ethanol Treatment on N-Methyl-D-aspartate R1 Splice Variants in Fetal Cortical Neurons*

Meena KumariDagger

From the Department of Pharmacology, University of Texas Health Science Center, San Antonio, Texas 78229-3900

Functional N-methyl-D-aspartate receptors consisting of NR1 and NR2 subunits are an important site of action of ethanol. Chronic ethanol treatment increases the NR1 polypeptide levels in vivo and in vitro. Chronic ethanol treatment in vitro does not significantly alter the NR1 mRNA levels, even though under similar culture conditions ethanol (50 mM, 5 days) enhances the half-life of NR1 mRNA in fetal cortical neurons. To address this phenomenon, we determined by reverse transcription-polymerase chain reaction and Western blotting whether ethanol (50 mM, 5 days) has a splice variant-specific effect on the expression of the NR1 subunit in mouse fetal cortical neurons. This report analyzes for the first time the distribution of all NR1 splice variants in these neurons. Our data indicate the presence of NR1-3a,b and NR1-4a,b splice variants in cortical neurons. Chronic ethanol treatment significantly decreased the mRNA levels of exon 5-containing NR1 splice variants (NR1-3b and NR1-4b) (-E5/+E5 = 4.6 in untreated neurons and 6.1 in ethanol-treated neurons) and had no effect on the mRNA levels of NR1-3 (+E21/-E22) and NR1-4 (-E21/-E22) splice variants. At the polypeptide level, chronic ethanol treatment significantly reduced exon 5-containing splice variants (NR1-3b and NR1-4b). However, ethanol (50 mM, 5 days) induced a significant increase in polypeptide levels of NR1-4 (-E21/-E22), without any effect on NR1-3 (+E21/-E22) polypeptide levels. These results demonstrate that chronic ethanol treatment has a selective effect on the expression of NR1 splice variants at both the mRNA and polypeptide levels in mouse fetal cortical neurons.

* This work was supported by National Institute on Alcohol Abuse and Alcoholism Grant AA12070.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Pharmacology, University of Texas Health Science Center, 7703 Floyd Curl Dr., Mail Code 7764, San Antonio, TX 78229-3900. Tel.: 210-567-4264; Fax: 210-567-4226; E-mail: kumari@uthscsa.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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