1. Acute Ethanol Exposure Alters the Metabolomic Profile in Rat Brain as Measured by High Resolution Magic Angle Spinning 1 H Magnetic Resonance Spectroscopy (HRMAS 1 H-MRS) Shonagh O’Leary-Moore Departments of Psychiatry and Psychology

2. Acute Alcohol Exposure-Neurobiology Enhances GABA-A activated Cl- Influx NMDA receptor antagonist Potentiates 5HT 3 receptors, CA +2 channel (EtOH actually probably inhibits most voltage gated ion channels) Stimulates DA release Differential susceptibility in various brain regions due to distribution of various receptor subtypes From: Oscar-Bermas & Marinovic, Alcohol Research and Health, 27, 125, 2003. Purpose: Determine the regional metabolic profile in rat brain following a binge exposure to alcohol using HR-MAS 1H MRS and HPLC.

3. Experimental Design & Methods n = 18, Male Sprague-Dawley Rats, ~250-300 g Subjects intubated with either 0 or 6 g/kg of 20% w/v of ethanol 1 st intubation (0 or 3 g/kg EtOH) 2 nd intubation (0 or 3 g/kg EtOH) 2 hr – Post EtOH and control blood and tissue collection 6 hr – Post EtOH blood and tissue collection BAC peak (225- 300 mg/dl) HIP VTA SN OCC CBV BS PST For HPLC HRMAS and HPLC

4. Spectral Processing - LCModel Quantification of Data and Custom Peak Fitting Algorithms for EtOH Comparison of Control and Ethanol- exposed spectra from the PST The presence of EtOH was clearly evident in all brain regions sampled

5. MDTOCCBSCBHIPPST 262626262626 Ala Asp Cre Cho GABA Gln Gly Glu GPC GSH Ins Lac NAA NAAG PEA PCh Tau RESULTS - Traditional Univariate Analyses of Neurochemical Data Neurochemical Profile of Acute Alcohol Exposure in all regions Neurochemical Profile of Acute Alcohol Exposure in the PST * * ** ** * * HVA/ DA ratios Acute EtOH increases DA turnover in the PST 2 hours after exposure Increase with alcohol Decrease with alcohol

6. PC1 accounted for ~21% of the variability in the data set and significantly distinguished control from alcohol exposed subjects (p < 0.05) RED – Negatively correlated with PC1 and decreased with acute EtOH exposure GREEN – Positively correlated with PC1 and increased with acute EtOH exposure MDTOCCBSCBHIPPST Cre-0.56-0.01-0.13-0.360.100.72 GABA-0.370.040.070.270.060.86 Gln-0.060.390.290.000.550.79 Gly-0.430.12-0.140.24-0.210.94 Glu-0.650.100.09-0.110.080.67 Ins-0.23-0.12-0.15-0.20-0.110.82 Lac-0.330.160.12-0.010.210.76 NAA-0.460.01-0.07-0.02-0.180.93 NAAG-0.38-0.06-0.08-0.090.590.31 PEA-0.22-0.12 -0.570.21 Tau-0.020.110.080.11-0.370.78 Principal Component Analyses PC1 Uses all of the variance from a data set Finds linear combinations of variables that account for maximum variance Components are uncorrelated (PC1 does not correlate with PC2)

7. Summary/Conclusions Acute EtOH alters the metabolic profile measured by HRMAS Brain EtOH levels can be seen and quantified by HRMAS EtOH exposure increased DA turnover, increased glutamine levels in the PST, BS, and HIP and decreased GABA in the MDT. Additional effects were seen in membrane phospholipids (PEA and GPC) PCA is useful to analyze HRMAS data Acknowledgements: Dr. Matthew Galloway Dr. John Hannigan Navid Seraji-Bozorgzad Kristen Prevost NIAAA F31 AA015224-01