(A) Schematic of the hippocampal granule cell layer demonstrating the sequence of preneuronal, early neuronal, and postmitotic cell types during postnatal neurogenesis (modified from ref. 13). (B–F) Qualitative images of Ki-67-IR cells that are type 1 (B), type 2a (C), type 2b (D), type 3 (E), and none of these cell types (F). Four panels for each cell type are shown indicating single-labeling of each marker, followed by a merge panel. Orientation of each of these panels is indicated by granule cell layer (GCL). Arrowhead in B points to a type 1 cell. Arrowhead in C points to a type 2a cell. Arrowhead in D points to a type 2b cell. Arrowhead in E points to a type 3, and in F, to a Ki-67-IR cell that was none of these cell types. Arrow in B points to a Ki-67-IR type 2a cell next to type 1 cell. [Scale bar in F merge, 10 μm (applies also to B–F).] (G) Quantitative data of calculated cell types that were type 1, type 2a, type 2b, type 3, or none of these cell types. Two separate analyses were performed to analyze significant differences between alcohol and control animals. First, unpaired t test revealed significant differences between control and alcohol animals with only type 1, type 2a, and type 2b cell types. Type 3 cells and cells not classified as any of the cell types (neither) were not significantly different from alcohol animals. Second, two-way ANOVA revealed a significant effect of alcohol on only type 2a cells. Data are expressed as mean ± SEM (n = 3–4 per group). *, P < 0.05, compared with controls by unpaired t test; #, P < 0.01, compared with controls by two-way ANOVA followed by Bonferroni's post hoc analyses.