Starting at 28 days of age (PND 28) P rats (n = 10) were given concurrent access to 15 and 30% ethanol for three 1-h sessions each day during the night cycle, as previously described (Bell et al., 2011). Sessions were conducted 5 days each week (no ethanol on weekends). Water and food were always available. Rats were killed by decapitation 3 h after the 1st ethanol access session on the 15th day of drinking (when rats were 49 days old). This 3-h time-point was selected in an attempt to maximize the response to alcohol on the expression of genes in tissue from rats that have a history of repeated adolescent binge drinking. A water control group (n = 10) was killed by decapitation at the same time. Brains were quickly removed and frozen in isopentane in dry ice. Brains were stored at −80° C until they were prepared for sectioning and micro-punching.
