Fig. 4.

Repeated CRF into the BLA, DRN, and d-BNST before ethanol diet reduces social interaction behaviors during withdrawal from chronic ethanol. The CRF dose (0.5 μg/0.5 μl) was microinjected twice at weekly intervals into the BLA, DRN, or d-BNST before exposure to 5 days of 4.5% ED (see Fig. 1 for protocol). A representation of the site and limits at which CRF was administered into each of these brain sites is presented in supplementary material. In the CD-vehicle and ED-vehicle groups, vehicle was administered into each of the brain sites (n = 3–4 for each site) and data for these vehicle injections were combined because a significant change across sites was not observed for these groups. When social interaction for the CD-vehicle group was compared with the ED-vehicle group, no significant effect was observed (P > 0.05). A group that received CRF and was on control diet only was not included for each of the present sites because previous data demonstrated that intracerebroventricular administration of CRF to rats that received control diet does not induce sensitization (Overstreet et al., 2004) and the repeated CRF in the CeA of control diet-treated animals likewise did not sensitize withdrawal-induced anxiety (Fig. 2). Social interaction was measured 5 to 6 h after the ethanol diet removal. The number of rats for each group is listed in Table 1, part 2. Representative sites where CRF was microinjected are presented in Supplemental Material. *, Significantly different from CD-vehicle and ED-vehicle [F(4,42) = 9.227, P < 0.001].