A Fatty Acid Oxidation-Dependent Metabolic Shift Regulates Adult Neural Stem Cell Activity.pdf

Spot14 driven cre recombinase s14icre knobloch et al

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Spot14-driven Cre recombinase (S14iCre) ( Knobloch et al., 2013 ) and yellow fluorescent protein Figure 3. Cpt1a Is Expressed in Adult NSPCs In Vivo (A) Immunohistological analysis of a reporter mouse expressing GFP under the Cpt1a promoter reveals high GFP expression in the SGZ of the DG, where NSPCs reside. Shown is a representative image of a brain section from a 2-month-old animal. (B) Co-stainings for Cpt1a-GFP and the prolif- eration marker Ki67 show that the majority of proliferating NSPCs do not express GFP, sup- porting the findings that FAO is specifically upregulated in quiescent NSPCs. Shown is a representative confocal image (maximum pro- jection) from a 2-month-old Cpt1a-GFP reporter mouse. Arrows indicate Ki67+/GFP ± cells; the arrowhead points to a Ki67+/GFP+ cell. The bar graph shows the percentage of GFP+ and GFP ± NSPCs out of all Ki67+ cells in the SGZ of the DG (mean ± SEM). (C) Cpt1a-GFP-positive NSPCs are almost all positive for the NSPC marker SOX2, and GFP+ processes co-stain with the NSPC marker Nestin. Shown is a representative confocal image (maximum projection) from a 2-month-old Cpt1a-GFP reporter mouse. Dotted lines show the outline of the granular zone of the DG, the boxed area is enlarged in the right panel. The bar graph shows the percentage of GFP+ and GFP ± NSPCs out of SOX2+ cells (mean ± SEM). (D) Cpt1a-GFP-positive NSPCs are almost all negative for the immature neuronal marker Doublecortin (DCX), suggesting that FAO is downregulated upon neuronal lineage commit- ment. Shown is a representative confocal image (maximum projection) from a 2-month-old Cpt1a-GFP reporter mouse. Dotted lines show the outline of the granular zone of the DG, the boxed area is enlarged in the right panel. The bar graph shows the percentage of GFP+ and GFP ± NSPCs out of DCX+ cells (mean ± SEM). Scale bars represent 200 m m (A), 50 m m (C and D, left), and 20 m m (B, C, and D, right). See also Figure S3 . 2148 Cell Reports 20 , 2144–2155, August 29, 2017
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(YFP) reporter alleles in the ROSA locus (R26YFP), inducing recombination at seven weeks of age. The low recombination efficiency of the S14iCre mouse line resulted in sparse labeling of cells, allowing the identification and classification of labeled progeny into potential clones according to their spatial clus- tering, similarly to previously published analyses referred to as clonal analysis ( Bonaguidi et al., 2011 ). However, it needs to be noted that the classification of cellular clusters into clones according to spatial distance only assumes common lineage and does not ultimately prove the presence of cells of clonal origin as this would require an additional level of genetic lineage tracing (e.g., through genetic bookmarking as previously used in NSPCs; Fuentealba et al., 2015 ). We first analyzed cell cluster size distribution 8 days after the first TAM administration using the sparse labeling method and found that size and composition of cellular clusters was not significantly altered between control and Cpt1a-cKO mice, although no larger clones were found in Cpt1a-cKO ( Figures S4 A and S4D). At this time point, slightly lower numbers of clus-
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  • Winter '19
  • Robert S Kiss
  • The Land, FAO, Neurogenesis, Fatty acid metabolism

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