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. 2021 Sep 20;4(11):e202000703. doi: 10.26508/lsa.202000703

Figure 2. Knock-down of choroid plexus App expression reduces cell proliferation in neurogenic niches.

Figure 2.

(A) Schematic of App ChPl knock-down model involving a single icv injection of AAV5 expressing shRNA targeting the mouse App sequence in wild type mice. Control (Ctrl) virus expresses a scrambled shRNA devoid of targets in mouse. (B) An icv injection of AAV5 leads to specific choroid plexus expression. Note the absence of eGFP in brain parenchyma. Scale bar, 200 μm. (C) Quantitative PCR analysis of App expression in the ChPl and Hc after ChPl App knock-down (n = 4 mice per group). (D) Western blot analysis of ChPl and Hc after ChPl App knock-down for quantification of APP protein levels normalized to actin, and assessing normalized APP metabolite ratios (n = 4 mice per group). (E, F) Analysis of cell proliferation in SVZ (E) and DG (F) by quantification of Ki67 positive cells after ChPl App knock-down (shRNA-App n = 7 mice, shRNA-Ctrl n = 5 mice). Arrows in (E) highlight regional differences in SVZ cells. Scale bars, 100 μm. **P < 0.01; ***P < 0.001; t test; all values, mean ± SD. ChPl, choroid plexus; Hc, hippocampus; SVZ, ventricular-subventricular zone; DG, dentate gyrus; icv, intracerebroventricular.