Fig 1: Histological analysis of embryonic wild-type, L1-deficient and reeler brain cortex sections and BrdU pulse chase labelling. (a) Nissl staining and layer-specific immunostaining of cortical sections of reeler, L1-deficient (L1 -/y) and wild-type (L1 +/y) mice at embryonic day 15; dashed lines indicate the area of the cortical plate (cp). Note that the L1 -/y cp appeared less compactly developed in comparison to the L1 +/y cp. The distribution of Tbr1-positive (red), Brn2-positive (red) and Cutl1-positive (green) cells in the L1 -/y cortex was slightly altered in comparison to the L1 +/y cerebral cortex, but these abnormalities were not as extensive as the misplacement of cells seen in the reeler cortex. Nuclear staining with DAPI (blue); mz, iz, svz: marginal, intermediate and subventricular zones; scale bars: 50 µm. (b,c) Altered distribution of BrdU-positive cells in the cerebral cortex of L1 -/y mice. A single BrdU dose (150 mg/kg) was injected intraperitoneally into timed-pregnant females carrying 13.5-day-old L1+/y and L1-/y embryos. Immunohistology for BrdU (red) was performed at birth (P0). The cerebral cortex was divided in 9 equidistant bins and the number of BrdU+ cells per mm2 was estimated for each bin (right); scale bars: 50 µm. Mean values ± SEM are shown for the numbers of BrdU-positive cells per mm2 analysing five sections from three animals per condition (*p < 0.05; Mann-Whitney U-Test).
Fig 2: Similar percentages of tdTomato-positive projection neurons are found in L2/3 and L5 of the ACC of WT and Cntnap2 KO mice.a Coronal section of the prefrontal cortex from a Grp:cre, tdTomato+ mouse at P14, highlighting the location of the ACC, and the position of TOM+ cells in the respective layers. b A representative coronal section shows staining of the ACC with an anti-CUX-1 antibody (for layer 2/3 projection neurons; green); TOM+ cells (red), and DAPI (all cell nuclei, blue). c The percentage of TOM+ cells, and of CUX-1+ cells, which colocalize with DAPI (all cells), is similar between WT and KO. d The percentage of TOM+ cells colocalising with CUX-1+ cells is similar between WT and KO mice. e Layer 5 projection neurons stained for CTIP2 (green), combined with DAPI staining (blue), and TOM+ cells (red). f Percentages of TOM+ and CTIP2+ cells colocalising with DAPI (all cells) are similar between WT and KO. g Roughly 9% of CTIP2-positive cells are TOM+ cells in both WT and KO. Percentages in charts represent means. Error bars represent SEM. WT: n = 4; KO: n = 4. CC corpus callosum. Scale bar in a = 100 μm, in b, c = 50 μm. Details of statistical analyses are shown in Supplementary Data 2.
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