(A) Representative traces of sIPSCs in CRF1+ neurons from male mice drinking water (left) or alcohol (right) during superfusion of Astressin 2B (200 nM, top) or Astressin 2B + CRF (200 nM;bottom). The traces shown are from the same neurons as depicted in Figure 7A. (B) Summary of the effects of Astressin 2B + CRF on sIPSC frequency (left) and effects of Astressin 2B + CRF normalized into percent change from baseline (right). Co-application of CRF + Astressin 2B following Astressin 2B pretreatment significantly increased sIPSC frequency in male CRF1+ neurons from water and alcohol drinking mice (***p < 0.001 by two-way RM ANOVA). When data were normalized to percent change from Astressin 2B alone, a significant difference between drinking groups was evident (*p < 0.05 by two-tailed t-test). CRF induced a significant increase in sIPSC frequency from baseline in the presence of Astressin 2B in CRF1+ neurons from water and alcohol drinking mice, but the increase was significantly greater in neurons from alcohol drinking mice (#p < 0.05 ##p < 0.01 by one-sample t-test). (C) Summary of the effects of Astressin 2B + CRF on sIPSC amplitude (left) and effects of Astressin 2B + CRF normalized into percent change from baseline (right). CRF + Astressin 2B significantly increased sIPSC amplitude in male CRF1+ neurons from both drinking groups (**p < 0.01 by two-way RM ANOVA). When data were normalized to percent change from baseline, CRF still stimulated an increase in sIPSC amplitude in the presence of Astressin 2B in CRF1+ neurons from both water drinking and alcohol drinking mice (#p < 0.05 by one-sample t-test). (D) Representative traces of sIPSCs in CRF1+ neurons from female mice drinking water (left) or alcohol (right) during superfusion of Astressin 2B (200 nM, top) or Astressin 2B + CRF (200 nM; bottom). The traces shown are from the same neurons as depicted in Figure 7D. (E) Summary of the effects of Astressin 2B + CRF on sIPSC frequency (left) and effects of Astressin 2B + CRF normalized into percent change from baseline (right). Co-application of CRF + Astressin 2B following Astressin 2B pretreatment significantly increased sIPSC frequency in female CRF1+ neurons from water and alcohol drinking mice (*p < 0.05 by two-way RM ANOVA). When data were normalized to percent change from baseline, CRF still induced a significant increase in sIPSC frequency from baseline in the presence of R121919 in CRF1+ neurons from alcohol drinking mice ( #p < 0.05 by one-sample t-test). The percent increase from baseline in water drinking mice was not statistically significant, but significant difference in effect size emerged between the two drinking groups. (F) Summary of the effects of Astressin 2B + CRF on sIPSC amplitude (left) and effects of Astressin 2B + CRF normalized into percent change from baseline (right). CRF + Astressin 2B did not alter sIPSC amplitude in female CRF1+ neurons from both drinking groups.