Fig. 5. The PVTBNST projection sex-dependently regulates binge alcohol drinking and avoidance behavior.
a Multiplexed chemogenetic approach for PVTBNST neurons. b Representative PVT image showing coexpression of the Gq-DREADD (mCherry) and Gi-KORD (mCitrine) in PVTBNST neurons. Scale bar = 50 μM. (Images were acquired for all mice except those used for slice electrophysiology recordings.) c–d Alcohol consumption during vehicle baseline vs. Day 4 ligand administration. c Clozapine n-Oxide (CNO; 5 mg/kg, i.p.; Gq-DREADD activation). 3xRM-ANOVA: no effects of sex, DREADD, CNO, or any interactions (Ps > 0.15; N’s = 9 CON M, 11 DREADD M, 10 CON F, 11 DREADD F). d Salvinorin B (SalB; 17 mg/kg, s.c.; Gi-KORD activation). 3xRM-ANOVA: sex × DREADD × SalB interaction (F1,26 = 8.76, **P = 0.007) and no main effects and no interactions (Ps > 0.05); post hoc two-tailed paired t-tests with Holm-Sidak corrections for effect of SalB: DREADD females (t26 = 3.08, *P = 0.019), all others (Ps > 0.65). N’s = 8 CON M, 8 DREADD M, 8 CON F, 6 DREADD F, with 0–3 mice/group excluded for below-criteria baseline drinking. e–g Effects of CNO on the elevated plus maze (EPM). N’s = 6 CON M, 8 DREADD M, 7 CON F, 7 DREADD F. e Percent time spent on the open arms. 2xANOVA: main effect of DREADD (F1,24 = 9.30, **P = 0.006) but no other effects (Ps > 0.45). Post hoc two-tailed unpaired t-tests with Holm-Sidak corrections within sex (Ps > 0.05). f Closed arm entries. 2xANOVA: no effects or interaction (Ps > 0.20). g Representative tracking heat maps. h–j Effect of SalB in open field test (OF). N’s = 4 CON M, 5 DREADD M, 4 CON F, 5 DREADD F. h, Percent time in center. 3xRM-ANOVA: main effect of time (F2,27.6 = 8.70, P = 0.003, not indicated) and no other effects or interactions (Ps > 0.10). i Distance traveled. 3xRM-ANOVA: main effect of time (F1.2,17.4 = 16.1, P = 0.0005, not indicated) but no other effects or interactions (Ps > 0.10). j Representative tracking heat maps. Data are presented as mean values ± SEM.