The present study investigates the effects of ethanol on neuronal and astroglial metabolism by using (1) H-[(13) C]-NMR spectroscopy in conjunction with infusion of [1,6-(13) C2 ]/[1-(13) C]glucose or [2-(13) C]acetate, respectively. A three compartment metabolic model was fitted to the (13) C turnover of GluC3 , GluC4 , GABAC 2 , GABAC 3 , AspC3 and GlnC4 from [1,6-(13) C2 ]glucose to determine the rates of TCA and neurotransmitter cycle associated with glutamatergic and GABAergic neurons. The ratio of neurotransmitter cycle to TCA cycle fluxes for glutamatergic and GABAegic neurons was obtained from the steady state [2-(13) C]acetate experiment and used as constraints during the metabolic model fitting. (1) H MRS measurement suggests that depletion of ethanol from cerebral cortex follows zero order kinetics with rate 0.18±0.04 μmol/g/min. Acute exposure of ethanol reduces the level of glutamate and aspartate in cortical region. GlnC4 labeling was found to be unchanged from a 15 minute infusion of [2-(13) C]acetate suggesting that acute ethanol exposure does not affect astroglial metabolism in naive mice. Rates of TCA and neurotransmitter cycle associated with glutamatergic and GABAergic neurons were found to be significantly reduced in cortical and subcortical regions. Acute exposure of ethanol perturbs the level of neurometabolites and decreases the excitatory and inhibitory activity differentially across the regions of brain. This article is protected by copyright. All rights reserved.