Figure 3.

MBD1-KO NSCs are impaired in transition to neuronal fate. A, Sample confocal images of Nes-GFP (green), DCX (red), and DAPI (blue) staining of brain sections from WT;Nes-GFP and MBD1-KO Nes-GFP mice. Scale bar, 20 μm B, Summary of quantification of Nes-GFP+ (Type 1 and Type 2a), Nes-GFP+DCX+ (Type 2b), and DCX+ (Type 3/immature neurons) in WT and KO mice. C–E, Quantification of individual cell types in the adult DG of WT and KO mice: C, GFP+DCX− cells, D, GFP+DCX+ and E, GFP−DCX+ (n = 7 per genotype), 2-way ANOVA, post hoc Bonferroni t test *p < 0.05, **p < 0.01. F, Schematic illustrations of retroviral expressing shMbd1 as well as GFP (Retro-shMbd1) injected into the adult DG. A timeline of the in vivo labeling of newborn neurons in the DG experiment and illustrations of vertical and parallel neurons is shown. G, Confocal images showing examples of retroviral-labeled (GFP+) and DCX+ (white) vertical and parallel neurons. Scale bar, 20 μm. H, I, Quantitative analysis showing that Retro-shMbd1-infection resulted in reduced differentiation into vertical neurons (H), but not parallel neurons (I). J, Sample confocal images of BrdU+ (red) cells in the SGZ identified with cell-type-specific markers: Nes-GFP+ (green) Type 2a cells (arrowhead, top), DCX and Nes-GFP double-positive Type 2b cells (asterisks, middle), DCX+ (white) Type 3 cells (arrows, bottom). Scale bar, 10 μm. K, Quantitative data showing the percentage of each cell type among total BrdU+ cells. After 24 h, MBD1-KO mice had significantly more BrdU+ cells that were Nes-GFP+ and significantly fewer were DCX+ compared with WT mice. L, Hypothetical model based on these results showing reduced transition of BrdU-labeled cells to Type 3 (DCX+) in MBD1-KO mice at 24 h after BrdU labeling. Data are presented as mean ± SEM, WT (n = 7), KO (n = 6), 2-way ANOVA, post hoc Bonferroni's t test, *p < 0.05, **p < 0.01.