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. Author manuscript; available in PMC: 2022 Sep 9.
Published in final edited form as: Mol Psychiatry. 2022 Mar 9;27(5):2502–2513. doi: 10.1038/s41380-022-01496-9

Figure 6. Chemogenetic inhibition of CeA CRF neurons does not reverse ethanol intake escalation or affective disturbance in alcohol-dependent mice.

Figure 6.

A-F. Crh-IRES-Cre male mice were injected with a Cre-dependent hM4Di-encoding vector in the anterior CeA. B. Firing rates were recorded from CeA mCherry+ and mCherry− neurons. Left: red fluorescence and differential interference contrast images of patched CeA neuron. Middle: representative traces before and during CNO application (500 nM). Right: CNO-induced change in firing rate (mCherry+, n=5; mCherry-, n=4). **, p<0.01, one-sample t-test. C. Experimental timeline for behavioral testing. Each box represents one week. AAV designates the timepoint at which the viral vector was injected, mice recovered for 3 weeks before resuming alcohol drinking sessions. The digging test (Dig) was performed 7 and 10 days after last vapor exposure (within-subject design). The novelty-suppressed feeding (NSF) test was conducted 14 days after last vapor exposure (between-subject design). D-F. Exposure to CIE increased ethanol intake (D), digging activity (E), and hyponeophagia (F). Acute CNO administration had no significant effect on these measures. Data were analyzed by two-way ANOVA (main effect of CIE: ##, p<0.01; ###, p<0.001). G. C57BL/6J mice subjected to the CIE-2BC procedure were injected with the CRF1 antagonist CP376395 30 min prior to 2BC. CP376395 reduced ethanol intake in both Air and CIE mice. Data were analyzed by two-way repeated-measures ANOVA (###, p<0.001, main effect of CIE; ***, p<0.001, Dunnett’s posthoc test). Data are shown as mean ± s.e.m. Number of mice per group is shown in the legend of each graph.