Figure 2
Loss of ephrin-A2 in apical progenitors results in reduced neuronal migration and the accumulation of progenitors in the ventricular zone. (a,b,c) Cytoarchitecture of the developing neocortex demarcated with the nuclear dye Hoechst at E14.5, E16.5 and E18.5 in wild type mice (a’) ephrin-A2 immunolabeling at E14.5, (a”) efnA2 messenger in situ hybridisation at E14.5 and (b’) at E16.5 reveals expression in the cells lining the ventricular surface of the lateral ventricle, in the MZ ((a’), arrowheads, *signal autofluorescent blood vessels) and in the cortical plate, double-labeled with NeuN ((b”) arrowheads). (c) Representative illustration of embryonic neocortex harvested at E18.5 stained with Hoechst following electroporation at E15.5 with (c’) GFP-tagged control scrambled shRNA. (c”) GFP-tagged construct coding for efnA2 for overexpression. (c”’) GFP-tagged shRNA to knockdown efnA2 (sh_efnA2-GFP) (d) ratio of GFP+ cells/compartment over the total number of GFP+ cells. (e) High magnification of the VZ of efnA2 knockdown electroporated at E15.5, analyzed at E18.5, sh_efnA2-GFP reporter (green), and Pax6 (magenta) (e’,e”) arrowheads signal electroporated cells also expressing the progenitor cell marker Pax6 (d) non-parametric Kruskal-Wallis test, error bar represent SEM CP cortical plate, IZ intermediate zone, MZ marginal zone, SVZ subventricular zone, VZ ventricular zone Scale bar (a”,b”’,c”’) 100 µm; (e”) 20 µm.
