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Department of Pharmacology, Louisiana State University Medical
Center, Shreveport, Louisiana 71130-3932 (L.J.C., G.S., D.N.),
Department of Physiology, University of Florida College of Medicine,
Gainesville, Florida 32610 (C.S.), and
Bowles Center for Alcohol
Studies, University of North Carolina at Chapel Hill, Chapel Hill,
North Carolina 27599 (F.T.C.)
The effects of prolonged ethanol exposure on excitatory amino acid
receptor stimulated nitric oxide (NO) formation were examined in
primary rat cortical neuronal cultures. Chronic ethanol (4 days, 100 mM) potentiated
N-methyl-D-aspartate (NMDA)-stimulated NO
formation as determined by measuring the conversion of
[3H]arginine to [3H]citrulline. In
contrast, chronic ethanol had no effect on NO formation stimulated
by kainate, 
-amino-3-hydroxy-5-methyl-4-isoxalonepropionic acid, or
the calcium ionophore ionomycin. Potassium chloride-stimulated NO
formation was also enhanced by chronic ethanol treatment, but this
effect was not seen in the presence of the ionotropic glutamate receptor antagonists MK-801 and 6-cyano-7-nitroquinoxaline-2,3-dione. Immunoblot analysis of expression of NR1, NR2A, and NR2B receptor subunits showed no difference between control and chronic
ethanol-treated cultures. In support of this apparent lack of change in
receptor density, there was no difference in the specific binding of
125I-MK-801 between control and chronic ethanol-treated
groups. These results demonstrate that prolonged ethanol exposure
selectively enhanced NMDA receptor-stimulated NO formation, which may
play an important role in alcohol dependence, withdrawal, and
alcohol-associated brain damage. These results also suggest that
chronic ethanol-induced increases in NMDA receptor function may not be
due to a simple increase in the number of NMDA receptors or change in
NMDA receptor subunit composition but may instead reflect more
complicated and subtle changes.
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