Figure 3.
SNAP23 deficiency enhances neural differentiation and apoptosis.(A and B) Tuj1 (A) or Tbr1 (B) stained in coronal sections of the cerebrum at E13.5. Scale bar, 50 µm. (C) Distribution of Tbr1-positive neurons in cerebral sections (n = 6 sections from three mice per genotype). (D) Density of Tbr1-positive neurons in cerebral sections (n = 6 sections from three mice per genotype). (E–G) Tuj1 (green in E), Calbindin (red in F, arrows), or Tbr1 (green in G, arrows) and DAPI (blue) stained in sagittal sections of the cerebellum at E13.5. The dashed lines and arrows in E indicate the ventricular surface and the abnormal localization of Tuj1-positive cells, respectively. Scale bars, 50 µm. (H) Number of Tbr1-positive neurons in cerebellar sections (n = 4 sections from three mice per genotype). (I) Ki67 (green) and BrdU (red) stained in coronal sections of the cerebrum at E13.5. The arrows indicate cells that exited the cell cycle (BrdU positive and Ki67 negative). Scale bar, 50 µm. (J) Quantification of the cell cycle exit rate in cerebral sections (n = 10 sections from three mice per genotype). The number of BrdU-positive/Ki67-negative cells was divided by the total number of BrdU-positive cells. (K) Ki67 (green) and BrdU (red) stained in sagittal sections of the cerebellum at E13.5. The arrows indicate cells that exited the cell cycle. Scale bar, 50 µm. (L) Quantification of the cell cycle exit rate in cerebellar sections (n = 3 sections from three mice per genotype). (M and N) Cleaved Caspase-3 stained in coronal sections of the cerebrum (M) and in sagittal sections of the cerebellum (N) at E13.5. Arrows indicate cleaved Caspase-3–positive cells. Scale bars, 50 µm. (O) Percentage of cleaved Caspase-3–positive cells in cerebral and cerebellar sections (n = 3 sections from three mice per genotype). (P and Q) TUNEL and Pax6 (P) or Tbr1 (Q) stained in coronal sections of the cerebrum at E14.5. Tbr1 staining overlaps with TUNEL-positive dying cells (arrows). Scale bars, 50 µm. The box plots represent the maximum and minimum values. The centerlines of the box plots represent the medians of the data. The bar graphs represent the means ± SEM. Significance was calculated by using two-tailed paired Student’s t tests. Statistical significance is indicated by *P < 0.05, **P < 0.01, or ***P < 0.001. Ctrl, control; EGL, external granular layer; NTZ, nuclear transitory zone; RL, rhombic lip; VZ, ventricular zone.

SNAP23 deficiency enhances neural differentiation and apoptosis. (A and B) Tuj1 (A) or Tbr1 (B) stained in coronal sections of the cerebrum at E13.5. Scale bar, 50 µm. (C) Distribution of Tbr1-positive neurons in cerebral sections (n = 6 sections from three mice per genotype). (D) Density of Tbr1-positive neurons in cerebral sections (n = 6 sections from three mice per genotype). (E–G) Tuj1 (green in E), Calbindin (red in F, arrows), or Tbr1 (green in G, arrows) and DAPI (blue) stained in sagittal sections of the cerebellum at E13.5. The dashed lines and arrows in E indicate the ventricular surface and the abnormal localization of Tuj1-positive cells, respectively. Scale bars, 50 µm. (H) Number of Tbr1-positive neurons in cerebellar sections (n = 4 sections from three mice per genotype). (I) Ki67 (green) and BrdU (red) stained in coronal sections of the cerebrum at E13.5. The arrows indicate cells that exited the cell cycle (BrdU positive and Ki67 negative). Scale bar, 50 µm. (J) Quantification of the cell cycle exit rate in cerebral sections (n = 10 sections from three mice per genotype). The number of BrdU-positive/Ki67-negative cells was divided by the total number of BrdU-positive cells. (K) Ki67 (green) and BrdU (red) stained in sagittal sections of the cerebellum at E13.5. The arrows indicate cells that exited the cell cycle. Scale bar, 50 µm. (L) Quantification of the cell cycle exit rate in cerebellar sections (n = 3 sections from three mice per genotype). (M and N) Cleaved Caspase-3 stained in coronal sections of the cerebrum (M) and in sagittal sections of the cerebellum (N) at E13.5. Arrows indicate cleaved Caspase-3–positive cells. Scale bars, 50 µm. (O) Percentage of cleaved Caspase-3–positive cells in cerebral and cerebellar sections (n = 3 sections from three mice per genotype). (P and Q) TUNEL and Pax6 (P) or Tbr1 (Q) stained in coronal sections of the cerebrum at E14.5. Tbr1 staining overlaps with TUNEL-positive dying cells (arrows). Scale bars, 50 µm. The box plots represent the maximum and minimum values. The centerlines of the box plots represent the medians of the data. The bar graphs represent the means ± SEM. Significance was calculated by using two-tailed paired Student’s t tests. Statistical significance is indicated by *P < 0.05, **P < 0.01, or ***P < 0.001. Ctrl, control; EGL, external granular layer; NTZ, nuclear transitory zone; RL, rhombic lip; VZ, ventricular zone.

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