Fig. 7: Downregulation of Cfl in DG decreases alcohol seeking during extinction and cue relapse, and increases alcohol consumption during relapse.

(A) The analysis of shCfl efficiency. Representative microphotographs of double immunolabeling with α-GFP and α-cofilin, combined with DAPI (nuclear marker) in the primary hippocampal cells culture, and summary of data showing cofilin levels in the neurons expressing shCfl (n = 21), or non-transduced neurons (n = 18) (Mann-Whitney U = 32). Cells were detected based on DAPI staining. (B) Experimental timelines and IntelliCage setups. Mice received dDG-targeted bilateral stereotactic injections of LVs encoding shCfl (n = 15) or shLuc (n = 14). Mice were trained to drink alcohol (4–12% and Free access) and AUD-related behaviors were tested: motivation to drink alcohol (M), alcohol seeking during withdrawal (W) and cue relapse (CR), alcohol drinking during alcohol relapse (AR), and alcohol seeking during a persistence test (P). (C, D) Summary of data showing individual scores in alcohol consumption during free access periods (C) and AUD-related behaviors (D). The animals were divided into two groups (bottom 70% and top 30% of the population) based on their performance in the tests. A significant effect of shCfl was observed on extinction of alcohol seeking during withdrawal (two-way ANOVA and LSD post hoc tests, effect of virus: F(1, 25) = 8.37, p = 0.008), alcohol seeking during cue relapse (F(1, 24) = 7.08, p = 0.014), and alcohol consumption during alcohol relapse (F (1, 25) = 6.87, p = 0.015). We found no effect of shCfl on motivation to alcohol (F(1, 25) = 0.141, p = 0.711), persistence in alcohol seeking (F(1, 25) = 0.299, p = 0.589) and alcohol consumption during free access periods (F(1, 25) = 3.30, p = 0.081). Data are presented as means +/- SEM.