Fig. 4: Age-dependent location defects of quiescent and activated neural stem cells and progeny in vivo.
a, Design of immunofluorescence experiments for quantifying the location of qNSCs/astrocytes and aNSCs in vivo in coronal brain sections. b, Schematic depicting how distance of cells to the ventricle were quantified. c, Representative images of immunofluorescence staining of coronal SVZ sections from young and old GFAP-GFP mice. The yellow box denotes the inset containing a qNSC/niche astrocyte (arrow) and an aNSC (arrowhead). The ventricular lining is indicated by a dashed white line (see Extended Data Fig. 9a for demarcation of ventricle wall with vinculin). Green, GFAP (astrocyte/NSC); pink, Ki67 (proliferation); blue, DAPI. Scale bar, 50 μm. d, NSC distance to the ventricle for qNSCs and niche astrocytes (Ast) and aNSCs in serial coronal sections (left) of young and old SVZs from mixed-sex GFAP-GFP mice and sagittal sections (right) of young and old SVZs from male C57BL/6 mice. Each dot represents the mean distance from the ventricle per mouse. Serial coronal sections: n = 4 young and n = 4 old mixed-sex GFAP-GFP mice, combined over four independent experiments. Sagittal sections: n = 5 young and n = 5 old male C57BL/6 mice, combined over two independent experiments. e, Design of immunofluorescence experiments to assess the location of EdU-labeled aNSCs/NPCs and neuroblasts in the SVZ neurogenic niche in vivo. i.p., intraperitoneal. f, Representative images of immunofluorescence staining of sagittal SVZ sections of young and old male C57BL/6 mice 4 h after EdU injection. Green, EdU; pink, Ki67 (aNSC/NPC/neuroblast); red, DCX (neuroblast); blue, DAPI. The dashed white line indicates the ventricle wall and arrows indicate EdU+ cells. Scale bar, 50 μm. g,h, Distance to the ventricle for EdU+ aNSCs/NPCs (g) and EdU+ neuroblasts (h) in sagittal sections of young and old SVZs 4 h after EdU injection. Each dot represents the mean distance from the ventricle per mouse. f–h, n = 5 young and n = 5 old male mice, combined over two experiments. All data are the mean ± s.e.m. All statistical comparisons were made using a two-tailed Mann–Whitney test. Figures in a, b and e were created with BioRender.com.
