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[Preprint]. 2024 Jan 14:2023.09.30.560339. Originally published 2023 Oct 2. [Version 2] doi: 10.1101/2023.09.30.560339

Figure 1. Experimental overview.

Figure 1.

(A) General timeline of the experiments. (B) Detailed timeline of the in vivo experiments: 9 daily alcohol injections, collections for blood alcohol levela (BAL) at intoxication and withdrawal, 1h and 24 h following injection, treatments on day 10, 24 h in withdrawal, with digging tests after 30 min and perfusion 90 min later. (C) BAL results for intoxication (1 h) and withdrawal (24 h). (D-F) Behavior data of animals following treatment with R121919 (purple), naltrexone (orange) or saline (black) treatment following chronic alcohol (or saline control, in lighter hues) for bouts of digging (D), length of digging (E), and number of marbles buried (F) (males = blue, females = pink). Analyzed by 2-way ANOVA, *p<0.05, **p<0.01. (G) Detailed timeline of brain immunolabeling and clearing. (H) Representative images of the brains in the last step of the protocol turning transparent through refraction index matching in dibenzylether. (I) Timeline of imaging and image processing with resampling and aligning FOS to autofluorescene to reference atlas.