Figure 5
Increase in neurogenesis in the subgranular zone (SGZ) following tdo deletion. (A) H&E staining of the dentate gyrus (DG) of paraffin-embedded coronal brain sections (5 μm) of 13-week-old Tdo+/+ and Tdo-/- mice. Center and right panels, higher magnification views. Right panels, PSA-NCAM-immunostaining. (B-I) Estimation of proliferating neural precursors and neurogenesis in the hippocampus of 13-week-old Tdo+/+ and Tdo-/- mice. (B) Total number of nuclei/section in the SGZ and granular cell layer (GCL) is shown as total cells. (C) BrdU/Ki67-double staining in the DG at 24 h after BrdU injection. (D-F) BrdU-, Ki67-, and double-labeled cells are indicated as a percentage of total cells. (G) Double-immunostaining of anti-BrdU and neural markers (Nestin, GFAP, PSA-NCAM, or DCX) in the SGZ of Tdo-/- mice at 24 h after BrdU injection. (H) BrdU-immunostaining in the DG at 28 days after BrdU injection. Nuclei were stained with TO-PRO-3 (blue). (I) TuJ1 and NeuN were co-labeled with anti-BrdU at 28 days after BrdU injection. Orthogonal images show three dimensional analyses of individual cells marked by intersecting lines in the x, y, and z axes. Bars: 100 μm (C, H) and 30 μm (G, I). Data represent means ± S.E. *, p < 0.05 versus Tdo+/+ mice.