Acetate Mediates Alcohol Excitotoxicity in Dopaminergic-like PC12 Cells

Neuronal excitotoxicity is the major cause of alcohol-related brain damage, yet the underlying mechanism remains poorly understood. Using dopaminergic like PC12 cells, we evaluated the effect of N-methyl-D-aspartate receptors (NMDAR) on acetate induced changes in PC12 cells: cell death, cytosolic calcium and expression levels of the pro-inflammatory cytokine, tumor necrosis factor alpha (TNFα). Treatment of PC12 cells with increasing concentrations of acetate for 4 hrs caused a dose-dependent increase in percent of cells staining positive for cell death using propidium iodide (PI) exclusion, and cytosolic reactive oxygen species (ROS) using Cell ROX detection, analyzed via flow cytometry. The EC50 value for acetate was calculated and found to be 4.40 mM for PI and 1.81 for ROS. Ethanol up to 100 mM had no apparent changes in the percent of cells staining positive for PI or ROS. Acetate (6 mM) treatment caused an increase in cytosolic calcium measured in real-time with Fluo-4AM which was abolished by co-ap...