Pharmacogenomic identification of small molecules for lineage specific manipulation of subventricular zone germinal activity
Fig 4
LY-294002 promotes dorsal subventricular zone (dSVZ)-oligodendrogenesis.
Pups were treated for 3 days with LY-294002 or saline/DMSO and examined by immunolabeling. (A, B) Periventricular sections show greater Olig2 immunostaining in LY-294002 in more dorsal periventricular regions compared to controls, illustrated in expanded insets. In the lateral SVZ, LY-294002 reduced Olig2 expression, as indicated by arrows (B). Arrowheads in (B) show reduced nuclei density in the dorsolateral horn of the SVZ where neuronal precursors (NPs) migrate. Scale bar in (A) = 200 μm. (C) Arrowheads show a loss of EdU in GFAP+ cells directly facing the lateral wall in LY-294002 compared to controls. Arrows show examples GFAP+ cells that have not incorporated EdU that were increased following LY-294002. (D) Single plane confocal micrographs show greater EdU+\Olig2 colocalization in LY-294002 and single panel captions of single planes illustrate that most newly generated Olig2+ cells co-express EdU and Ascl1 (arrows) or have absent or lower levels of Ascl1 (arrowheads). Scale bar in (D) = 10 μm in captions of (D), 15 μm in main panels of (D), 15 μm in (C), and 10 μm in (D). (E) Confocal micrographs illustrate a lower density of Dcx+ cells in LY-294002 and a loss of their proliferative status (compare Dcx+ cells with arrowheads to those marked by arrows). (F-H) Quantification of changes in GFAP+/EDU+ or EdU- cells directly facing the wall of the lateral ventricle (F). Quantification of changes in EdU+ cells expressing progenitor markers (Dcx, Olig2, or none of these markers [NP-]) (G). Quantification of changes in Olig2+/Ascl1+ or Ascl1- cells (H). Data are mean ± standard error of the mean (SEM) normalized to controls (n = 5 for control and LY-294004 for all quantifications); significance was tested using unpaired t test throughout versus respective control; **p < 0.01; ***p < 0.001.