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. 2013 Oct 30;33(44):17490–17505. doi: 10.1523/JNEUROSCI.2042-13.2013

Figure 3.

Figure 3.

Inactivation of Smo in mGfap-Cre-expressing cells leads to a gradual reduction in neurogenesis. A, B, Cresyl violet staining of a cross section through the RMS of P15 mGfap-Smo CKOs shows no drastic change in the RMS size from that in control animals (Smolox/lox or Smolox/+). C–H, Cross-section of the RMS in adult mGfap-Smo CKOs and controls (Smolox/lox) reveals a progressive reduction of the RMS in controls but a substantial loss of DCX+ migrating progenitors by 1 year of age in mutant animals. Dashed line in G and H indicates expected location of RMS. Scale bar: (in A) A–H, 100 μm. I, J, Quantitative analysis of proliferating KI67+ progenitors and BrdU+ LRCs at P15 (I) and in 3-month-old mGfap-Smo CKOs and controls (Smo ctrl; J) demonstrates a dramatic depletion of the SVZ LRC pool and a concurrent reduction in KI67+ progenitor cell number at 3 months. K, By 1 year of age, the level of neurogenesis appears to decrease in control animals and SVZ proliferation in mGfap-Smo CKOs continues to be significantly lower than in controls and appears lower than at 3 months (*p < 0.05; ***p = 0.0001 unpaired t test; mean ± SEM). L, GFAP and S100β marker analysis of the LRCs in the SVZ of controls and mGfap-Smo CKOs reveals a significant decrease in GFAP+ S100β− LRCs (green) and an apparent complementary increase in the proportion of GFAP− S100β+ cells (yellow; *p < 0.05; ***p = 0.0001; ****p < 0.0001, 2-way ANOVA with Sidak's post hoc test; mean ± SEM). M, Normalizing the number of each subpopulation of LRCs to the number of cells per section reveals a specific loss of GFAP+ S100β− NSCs (green) but no increase in the number of GFAP− S100β+ glial/ependymal cells (yellow) or GFAP+ S100β+ astrocytes (red). As expected for cells that originate from NSCs, the number of GFAP− S100β− cells is significantly decreased (blue), and is reflected by a decrease in the number of OLIG+ LRCs (blue vertical stripes) and PSA-NCAM+ LRCs (blue horizontal stripes) in the SVZ of mGfap-Smo CKOs compared with controls (****p = 0.0001 and ***p = 0.0007, 2-way ANOVA with Sidak's post hoc test; mean ± SEM).