Figure 7.

Radial migration of early (E13.5) and later (E15.5) born neurons in the neocortical regions with BrdU birth dating. A, BrdU immunofluorescence staining of E17 cortex (BrdU administrated to pregnant Manf^+/- mice at E13.5). B, The number of BrdU-labeled cells in each layer was counted and shown as percentages of total labeled cells number in the neocortex. The relative number of BrdU-positive cells were found to be decreased in CP and SP of Manf^-/- mice compared to WT littermates, and increased levels in VZ/SVZ (n = 3; **p < 0.01, ***p< 0.001). C, BrdU was administered at E15.5 and the distribution of the labeled cells was examined at E19. In the WT cortex, increased number of labeled cells (green) were found in in the Layers II/III compared to that in Manf^-/- cortex. In contrast, increased number of BrdU-positive cells were found in the cortical Layers IV and V/VI in the Manf^-/- mice compared to the WT mice. D, To quantify the distribution of BrdU-positive nuclei, the cerebral cortex was divided into five area from the pial side to the ventricular side. The number of labeled cells in each bin was counted and shown as percentages of total labeled cells number in the CP (n = 3, *p < 0.05, ***p < 0.001 for Manf^-/- mice compared to WT). Nuclei were counterstained with DAPI (blue). E, F, Immunofluorescent staining of coronal sections of neocortex with anti-Nestin and anti-Reelin antibodies. Radial glial architecture and Reeling expression in were similar in WT and in mutants; scale bars: 50 µm (A, C) and 25 µm (E, F).