9 and 10. Rats were killed by decapitation 3 hours after the 4th session at the end of the second 2-week relapse period. This 3-hour time point was selected in an attempt to maximize the response to alcohol on the expression of genes in tissue from rats that have had a history of repeated excessive binge drinking. The brains were quickly removed and frozen in isopentane on solid frozen CO2. Age-matched alcohol-naïve water-control female P rats (n = 8) were killed and brains were removed in a similar manner. Brains were stored at −70°C until sectioned.
