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. 2013 May 29;33(22):9408–9419. doi: 10.1523/JNEUROSCI.2700-12.2013

Figure 9.

Figure 9.

IGF2 stimulated proliferation of Dgcr8+/− neural progenitor cells both in vitro and in vivo and rectified the hippocampus-dependent spatial working memory deficits in Dgcr8+/− mice. A, Bright-field image of neurospheres cultured in the indicated conditions. Neurospheres derived from Dgcr8+/+ and Dgcr8+/− mice hippocampus were cultured in serum-free neurosphere medium in the presence or absence of IGF2. Note that the size of neurosphere was increased by IGF2 stimulation. Scale bar, 100 μm. B, Quantification of the number of sphere-forming progenitors in each condition (n = 3 each). Note that the number of neurosphere was increased by IGF2 stimulation (*p < 0.05, Dgcr8+/+ + EGF + bFGF + IGF2 vs Dgcr8+/+ + EGF + bFGF, two-tailed t test, n = 3, mean ± SEM; *p < 0.05, Dgcr8+/− + EGF + bFGF + IGF2 vs Dgcr8+/− + EGF + bFGF, two-tailed t test, n = 3, mean ± SEM). C, E, G, Immunolocalization of proliferating neural progenitor cells, immature neurons, and terminally differentiated neurons in the dentate gyrus after intrahippocampal administration of PBS or IGF2. Three days after injection, brains were harvested and then brain sections were stained with anti-SOX2 (green), anti-Ki67 (red), and anti-DCX (green) antibodies (C, E). For terminally differentiated neurons, brains were harvested at 28 d after injection and then stained with anti-BrdU (red) and anti-NeuN (green) antibodies (G). The number of SOX2/Ki67-, DCX-, and BrdU/NeuN-positive cells in the Dgcr8+/− brain was increased (C, E, G). The nuclei were stained with DAPI (blue). The second and fourth columns from the left show magnified views of the boxed regions. Scale bar, 50 μm. D, F, H, Quantification of the number of SOX2/Ki67-, DCX-, and NeuN/BrdU-positive cells in the dentate gyrus. For quantitation, immunopositive cells were counted over five unbiased images at 20× magnification per animal in three animals for each genotype. Note that the number of SOX2/Ki67-, DCX-, and NeuN/BrdU-positive cells in Dgcr8+/− was increased by IGF2 administration (*p < 0.05, Dgcr8+/− + IGF2 vs Dgcr8+/− + PBS, two-tailed t test, n = 3, mean ± SEM).