FIGURE 1.

Experiment design. (A) Timeline refers to behavioral analysis and blood ethanol concentration (BEC) in non-stressed controls and socially defeated mice. On day 1, mice were exposed to sweet solution two-bottle choice for 24 h (habituation). On days 2–4 (baseline intake), 5, 14, and 21 mice were offered sweet solution in the two-bottle choice procedure for 90 min. On days 5–14, stressed mice were weighed and socially defeated. Controls were weighed daily. On day 22, controls and socially defeated mice were tested in the elevated plus-maze (EPM), and next day in the tail suspension test (TST). On day 24, mice were given continuous access to ethanol and water for 4 weeks before the surgical procedure to implant cannulae into the bed nucleus of the stria terminalis (BNST). After recovery, mice were infused with selective CRF receptors antagonists (CP376,395 or astressin2B) and tested for alcohol drinking. Each mouse received three microinjections, with a 48-h interval between them, in a design that counterbalanced saline and drug treatments. On day 66, mice were euthanized and had the brains and blood samples collected for histological and BEC analysis. (B) Timeline refers to analysis of CRF, CRFR1 and CRFR2 mRNA expression in the BNST in non-stressed controls and socially defeated mice. On days 1–10, controls were weighed daily and stressed mice were weighed and socially defeated. On day 11, mice were euthanized and had the brains collected for mRNA expression analysis.