Fig. 3: Cfl is upregulated in DG of the ≥ 2 crit animals during extinction of alcohol seeking.
From: Hippocampal Cofilin and CFL1 gene variants are linked to Alcohol Use Disorder phenotypes
(A, B) Experimental timeline. Mice were trained to drink alcohol (n = 26) or water (alcohol-naive, n = 8) and AUD-related behaviors were tested. (B) AUD scores were calculated to identify the < 2 crit vs ≥ 2 crit drinkers. < 2 crit vs. ≥ 2 crit mice significantly differed in alcohol seeking during withdrawal (Mann Whitney test, U = 2). Mice were sacrificed and perfused after free alcohol access period (day 83) or alcohol withdrawal (day 90). The brains were cut and brain sections immunostained to detect cofilin (Cfl). (C) Schematic representation of the analyzed dDG regions. GCL, granule cell layer; PoDG, polymorphic layer of DG; ML, molecular layer. (D) The analysis of Cfl fluorescent immunostaining. Representative microphotographs and summary of data (repeated measures three-way ANOVA with Šídák’s multiple comparisons test, effect of region: F(1,40, 30,8) = 16.6, p < 0.001; effect of phenotype: F(1, 23) = 0.169, p = 0.685; effect of time: F(1, 23) = 7.33, p = 0.013; effect of region × AUD: F(2, 44) = 0.0189, p = 0.981; effect of region × time: F(2, 44) = 3.36, p = 0.044; effect of phenotype × time: F(1, 23) = 14.2, p = 0.001. (E) The analysis of Cfl colocalization with presynaptic marker, synaptotagmin 1 (Syt1). Representative microphotographs and summary of data (repeated measures three-way ANOVA with Šídák’s multiple comparisons test, effect of region: F(1.71, 34.3) = 26.3, p < 0.001; effect of phenotype: F(1, 22) = 0.598, p = 0.447; effect of time: F(1, 22) = 1.88, p = 0.184; effect of region × phenotype: F(2, 40) = 0.919, p = 0.184; effect of region × time: F(2, 40) = 4.04, p = 0.025; effect of phenotype × time: F(1, 22) = 11.8, p = 0.002. (F) The analysis of Cfl colocalization with a postsynaptic marker, PSD-95. Representative microphotographs and summary of data (repeated measures three-way ANOVA, effect of region: F(1.84, 40.5) = 4.59, p = 0.018; effect of phenotype: F(1, 22) = 0.0296, p = 0.865; effect of time: F(1, 22) = 1.34, p = 0.259; effect of region × phenotype F(2, 44) = 0.836, p = 0.440; effect of region × time: F(2, 44) = 1.27, p = 0.292; effect of phenotype × time: F(1, 22) = 1.06, p = 0.315. Mean ± SEM are shown. For B, D-F each dot on the graphs represents one animal.
