The study protocol was approved by the Institutional Animal Use and Care Committee of the University of Massachusetts Medical School. All the methods were carried out in accordance with the approved guidelines. CX3CR1eGFP/eGFP mice were provided by Dr. Dorothy Schafer from the University of Massachusetts Department of Neurobiology. CCR2RFP/RFP mice were purchased from the Jackson Labs. CX3CR1eGFP/wt CCR2RFP/wt mice were bred in-house. Wild-type mice were purchased from the Jackson Labs and allowed to acclimate to our animal medicine facility for at least 1 week prior to experimental use. Eight-week-old female mice were used for alcohol feeding experiments. Eight-week-old C57BL/6J mice were divided into alcohol- and pair-fed (control) groups as well as two CVC alcohol-fed groups. Alcohol-fed mice received 5% (v/v) alcohol in Lieber-DeCarli liquid diet ad libitum as previously described [17]. Pair-fed animals received a calorie-matched liquid diet. Mice were provided continuous access to alcohol or calorie-matched liquid diet until the time of anesthesia and were immediately transcardially perfused once appropriate sedation was achieved.
