the effects of alcohol on cell death, adjacent tissue sections were dyed for a common marker of cell death, FJB, and FJB-positive (FJB+) cells were counted in the dentate gyrus and subgranular zone of the hippocampus. As shown in Figure 7, FJB + cells were found throughout the dentate gyrus of the alcohol treated sections, and intriguingly most were along the subgranular zone, where proliferation occurs. Indeed, the number of FJB+ cells was significantly increased (p<0.05) in the ethanol-exposed subjects (n=6), as compared to controls (n=3). Capitalizing on the neutral red counterstain of the Ki-67 labeled sections, we then confirmed the cell death observed with FJB staining, by quantifying the number of pyknotic cells observed in the dentate gyrus. Similar to FJB, pyknotic cells were located sparsely throughout the granule cell layer but the majority were located along the subgranular zone. As shown in Figure 7, there was a significant increase (p<0.01) in the number of pyknotic cells in the tissue of the ethanol treated subjects (n=6) compared to controls (n=6). Thus, cell death is observed in the dentate gyrus immediately following binge alcohol exposure in adolescence. Previous work has suggested that binge alcohol exposure in adult rats results in
