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. 2018 Sep 26;38(39):8496–8513. doi: 10.1523/JNEUROSCI.0632-18.2018

Figure 3.

Figure 3.

Impairment of maturation of newborn granule cells following CSPG digestion. A, Representative reconstruction images showing Sox2−/DCX+/CR+/S100β− newborn granule cells in the dentate gyrus of mice treated with P-ase (left) or chABC (right). B–E, Morphometric data obtained from 3D reconstruction of the dendrites and cell bodies of newborn granule cells: branch numbers (B; F(3,12) = 0.6292, p = 0.6100), total dendritic length (C; F(3,12) = 24.4539, p = 0.0002), second-order dendritic length (D; F(3,12) = 0.0339, p = 0.9912), and third-order dendritic length (E; F(3,12) = 5.5738, p = 0.0125). F, Sholl analysis of the dendritic morphologies of newborn granule cells. The numbers of intersections at 70 μm (F(3,12) = 7.3223, p = 0.0048), 80 μm (F(3,12) = 23.9210, p < 0.0001), 90 μm (F(3,12) = 22.8820, p < 0.0001), 100 μm (F(3,12) = 10.8076, p = 0.0010), and 110 μm (F(3,12) = 6.9502, p = 0.00577) from the cell body are lower in mice treated with chABC or chAC than in mice treated with P-ase. G–J, Based on the fluorescent quadruple staining for DCX (green), CR (magenta), BrdU (cyan), and NeuN (yellow), DCX−/CR−/BrdU+/NeuN+ mature granule cells (white arrowheads) are defined in the dentate gyrus of mice treated with P-ase (G), chABC (H), chAC (I), or chB (J). K, Schematic diagram of the experiment. One week after intraperitoneal injection of BrdU, mice were treated with P-ase, chABC, chAC, or chB and killed 3 weeks later. L, NDs of BrdU+ mature granule cells in the dentate gyrus of mice treated with different enzymatic reagents (F(3,12) = 40.6887, p < 0.0001). M, N, Fluorescent staining for Casp-3 (magenta) and DAPI (cyan) in the dentate gyrus of mice treated with P-ase (M) or chABC (N). O, NDs of Casp-3+ apoptotic cells in the dentate gyrus of mice treated with different enzymatic reagents (F(3,12) = 2.4941, p = 0.1097). Data were analyzed by one-way ANOVA with post hoc Tukey's HSD test and are shown as mean ± SD (n = 4 mice in each group). Statistical significance: *p < 0.05, **p < 0.01 (vs. P-ase). Scale bars: A, 30 μm; N, 30 μm (applies to G–J, M, and N).