Figure 2

Fluoro-Jade B staining in the cortex and dentate gyrus of C57BL/6 mice. Brain sections from C57BL/6 mice treated with either water or ethanol daily for 10 days were double-stained with Neu-N antibody and Fluoro-Jade B as described in Materials and Methods. Neu-N immunolabeling was visualized by using Alexa Fluor 555. (A) Fluorescent intensity of Fluoro-Jade B positive cells in cortex and dentate gyrus was quantified by BioQuant image analysis system. The cortex and DG of chronic ethanol-treated mice had more Fluoro-Jade B positive cells than water control mice 24 hrs after the last dose of ethanol treatment. These cells are colocalized with Neu-N, a neuronal marker, suggesting neuronal death. (B) Representative images for Fluoro-Jade B (green), Neu-N (red) and colabeling of Fluoro-Jade B and Neu-N (yellow) from cortex and dentate gyrus in ethanol-treated mice. ** P < 0.01, compared with the water control mice (n = 10). Scale bar = 30 μm; inset 5 μm.