Figure 3

Immunohistochemical evaluation of the alcohol-induced inflammatory response. (A to F). TNF-α reactivity in the neural compartment in the CeA (A to C) and DVC (D to F) in control animals (A,  D; N = 4) and following chronic alcohol exposure (B,  E; N = 2) and 48 hours into withdrawal (C,  F; N = 2). Neurons are stained red with neuronal nuclear antigen (NeuN) and nuclei with DAPI shown in blue. The arrows in panels C., E. and F. show the expression of TNF-α (green) surrounding cells expressing the neural marker NeuN. Dual labeling of a cell for both NeuN and TNF is shown in yellow. (G to L). ICAM-1 reactivity (green) in the endothelial compartment in the CeA (G to I) and DVC (J to L), following chronic alcohol exposure (H, K) and 48 hours into withdrawal (I, L). Endothelia are stained red with RECA-1 and nuclei with DAPI shown in blue. The arrow in panel H shows the expression of ICAM-1 by cells also expressing the endothelial cell marker RECA-1. Panels I and L have arrows showing small cells with a high degree of ICAM-1 staining found commonly in the chronic and withdrawal tissue that are absent in the control condition. Dual labeling of a cell for both RECA-1 and ICAM-1 is shown in yellow. CeA, central nucleus of the amygdale; DAPI, 5 μg/ml 4′-6-diamidino-2-phenylindole; DVC, dorsal vagal complex; ICAM‐1, intercellular adhesion molecule 1;NeuN, neuronal nuclear antigen; RECA‐1, rat endothelial cell antigen‐1.