Binge-, chronic- and withdrawal-like ethanol conditions were used to study effects of ethanol exposure on MeCP2 regulation in forebrain-derived neural stem cells (NSCs). To study the methylation effects of alcoholic-like ethanol concentrations (70 mM; 320 mg/dL), cells were treated during the onset of differentiation for 48 hours (as a model of a binge effect) or 8 days (to mimic a chronic exposure). Additionally, withdrawal effects were studied 6 days after withdrawal of ethanol. In previous work, an altered cellular morphology was observed in ethanol-exposed, cultured glia and neurons, thus the morphological state of NSC was measured in response to ethanol treatment. This was assessed through the expression of neural and glial markers as well as measurement of neurite outgrowth and glial size. There were no significant changes in the numbers of cell-type specific markers. However, secondary and tertiary neurite outgrowth was increased in the chronic and withdrawal conditions. Glial size was also affected in that a 1.5-fold increase was observed during chronic ethanol exposure, but there was no effect in the withdrawal condition. MeCP2 expression levels were upregulated during chronic and binge exposure (1.46-fold [p>0.05], 1.55-fold [p>0.001], respectively) while the withdrawal condition had a 1.38-fold reduction in MeCP2 transcripts.
