Figure 4.

Voluntary Running Restores Adult Neurogenesis and Cognitive Deficiencies of VPA-Treated Mice

(A) Representative images of brain sections including the hippocampal DG stained for BrdU (green) and with Hoechst 33258 (blue), 1 day after the last BrdU injection. See Figure 2A for the experimental timeline.

(B) Quantification of BrdU⁺ in the granule cell layer (GCL), 1 day (1d; n = 8 for each group) and 4 weeks (4w; n = 8 for each group) after the last BrdU injection, shows an increased number of BrdU⁺ cells in the hippocampus after voluntary running.

(C and D) Voluntary running recovers the proportion of BrdU⁺ cells that had differentiated into NEUN⁺ neurons (C) and the ones that still expressed SOX2 (D) among total BrdU⁺ cells at 4 weeks after the last BrdU injection (n = 4 for each group).

(E) Reduction of correct-arm alternation in Y-maze tests of VPA-treated mice is recovered by voluntary running (n = 7 for MC and VPA + RW; n = 8 for MC + RW and VPA).

(F–H) Voluntary running recovers the freezing response in conditioning (F; day 1) and in cued fear associative tests (H; day 3), but not in contextual fear associative tests (G; day 2; n = 7 for MC and VPA + RW; n = 8 for MC + RW and VPA). See also Figures S4B, S4D, and S4F for the time course of the freezing response and Table S2 for a summary of behavior data.

MC, prenatal methylcellulose (vehicle); MC + RW, prenatal methylcellulose and postnatal running; VPA, prenatal valproic acid; VPA + RW, prenatal valproic acid and postnatal running. Data are represented as means. Error bars indicate the SD. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, n.s., not significantly different, two-tailed t test. Scale bar, 100 μm. See also Figure S4 and Table S2.