CD68 is a lysosomal protein that is often used as a marker of phagocytosis in macrophages and particularly microglia [23, 24]. We hypothesized that ethanol consumption not only alters microglia morphology but also its phagocytic activity in the hippocampus. Microglial CD68 levels were measured by immunofluorescence, which demonstrated a decrease in the hippocampus (including CA1, CA3, and dentate gyrus (DG) regions) of alcohol-fed compared to pair-fed mice (Fig. 6a, b). This observation was corroborated by flow cytometric analysis of isolated microglia in which we also observed a decrease in microglial CD68 positivity in alcohol-fed mice (Fig. 6c). Interestingly, treatment with CVC did not affect microglial CD68 expression (measured as colocalization of CD68 and the microglial marker IBA1) except in the CA3 region of the hippocampus where CVC decreased CD68 expression (Fig. 6b). However, flow cytometry measurements of CD68+ microglia revealed an effect of CVC treatment as both inhibitor treatment paradigms increased CD68 positivity (Fig. 6c). Importantly, the flow cytometry is based on microglia from the total brain, whereas the immunofluorescence measurements were performed in the hippocampus only. Taken together, these
