Studies were conducted in adult drug-naïve C57BL/6J (B6) female mice from a colony maintained at the University of Texas. C57BL/6J (B6) female mice were utilized, as they are known to voluntarily consume more ethanol than males[28]–[30]. Original breeding pairs were purchased from Jackson Laboratories (Bar Harbor, ME) and mated at 8 weeks of age. Mice were initially group-housed in standard polycarbonate shoebox cages (four-five per cage), then moved to individual cages and allowed to acclimate for 1 week prior to treatment. DID animals were given an additional two-week acclimation period to adjust to the reverse light cycle (see below). Food (Prolab RMH 1800 5LL2 chow, TestDiet, Richmond, IN) and water were provided ad libitum, except as noted below. The colony rooms and testing rooms were maintained at an ambient temperature of 21±1°C, humidity (40–60%), and centrally controlled ventilation (12–15 cycles/h with 100% exhaust). All treatment and control groups consisted of 10 animals, except the treated CI group, which contained 11 animals. A 20% (v/v) ethanol solution was used for all studies. Ethanol and water bottles were weighed daily and animals were weighed every four days.
