Chronic ethanol inhalation procedures, tissue harvest, and brain dissection methods were as previously described [26–29]. Briefly, chronic intermittent ethanol vapor exposure (or air) was delivered in Plexiglas inhalation chambers to drug-naïve C57BL/6J (B6) male mice (8 treated and 8 controls per group). Ethanol treatments were performed in the laboratory of Dr. H.C. Becker (Medical University of South Carolina, Charleston, SC, USA). Mice were administered alcohol (1.6 g/kg; i.p.) and the alcohol dehydrogenase inhibitor pyrazole (1 mmol/kg; i.p.) prior to vapor ethanol exposure in inhalation chambers. Control subjects received pyrazole injections in saline and received similar daily handling. Chamber ethanol concentrations were monitored daily to induce stable blood ethanol concentrations within the range of 180–200 mg/dl. Ethanol was administered 16 h/day in 4 weekly cycles and alternated with 1 week in between cycles in which the mice were left undisturbed (mimicking drinking weeks). Animals were sacrificed at 3 time points: 0-, 8- and 120-hours following the last ethanol vapor or air treatment. RNA purification, quantification and quality assessment were performed as in [26]. Briefly, total RNAs were purified using the MagMax
