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. 2018 Dec 21;12:500. doi: 10.3389/fncel.2018.00500

Figure 3.

Figure 3

Aberrant cerebellar cortical development in VPA-treated mice during the early postnatal phase. (A) Timelines for the EdU labeling and sampling schedule. (B,C) Representative images for the EdU+ cells (red) with DAPI (blue) staining in sagittal sections of the cerebellum at P7 (B) and P10 (C). (D,E) Quantitative analysis for the density of the EdU+ cells (per 10,000 μm2) in the external granular layer (EGL) and internal granular layer (IGL) on P7 (D) and P10 (E). VPA-treated mice showed more EdU+ cells in P7 IGL (Mann Whitney test, U = 104, P = 0.0191) and fewer cells in P10 EGL (2-sample student t-test, t(53) = 3.416, P = 0.0012), suggesting a pre-onset of the granular cell (GC) precursors migration. N = 25 slices from six mice in the saline-treated group, and 15 slices from five mice in the VPA-treated group. (F) Representative images for terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) staining to label apoptotic cells (green) at P9. (G) Quantitative analysis of apoptotic cells. VPA-treated mice showed more TUNEL+ cells in the EGL (t(16) = 2.673 P = 0.0167) but less apoptosis in the IGL (t(16) = 3.75). N = 8 slices from three mice in the saline-treated group, and 10 slices from four mice in the VPA-treated group. ns, no significant difference; *P < 0.05; **P < 0.01. Scale bar, 100 μm in (B,C,F).