Pharmacogenomic identification of small molecules for lineage specific manipulation of subventricular zone germinal activity
Fig 6
Pharmacological stimulation of Wnt/β-catenin signaling rescues oligodendrocyte precursor (OP) and glutamatergic neuron progenitor numbers in the adult mouse.
(A) Quantitative PCR (qPCR) analysis reveals a pronounced decrease of Wnt targets genes Lef1 and Axin2 and pallial Emx1 and Tbr2 transcripts expression in the dorsal subventricular zone (dSVZ) between P6 and P60 (n = 3 for P6 and P60). Results are expressed as a percentage and normalized in comparison with Gapdh level of expression and compared using unpaired t test. (B) Representative coronal sections illustrating the pronounced and rapid decrease of Wnt canonical signaling in the βGal reporter mouse (βGAL+) and the parallel decrease of glutamatergic NPs (Tbr2+) and OPs (Olig2+) in the SVZ of mouse brain at the age of 6 d (P6), 2 mo (P60), and 4 mo (P120) (n = 3 individual animals for each time point). (C) Quantification of the average number of βGAL+, Tbr2+, and Olig2+ cells in the dorsal wall of the SVZ in P6, P60, and P120 mice (3 animals per age). (D) Representative pictures of Ki67, Tbr2, and Olig2 expression in the adult (P90) SVZ before and after treatment with GSK3β inhibitors (AR-A014418, not shown, and CHIR99021, shown). (E) Percentage increase of proliferation (Ki67, EdU), OPs (Olig2), glutamatergic NPs (Tbr2), and NSC (Mcm2/GFAP) numbers following intraventricular infusion of AR-A014418 (3–10 μM) and CHIR99021 (3–10 μM). Values are normalized compared to the controls (n = 5 for each of control, AR-A014418, and CHIR99021). Error bars represent standard error of the mean (SEM). ***, p < 0.001; **, p < 0.01; *, p < 0.05; t test. Scale Bar = 1 mm (B) and 50 μm (D).