Fig. 1: Altered laminar organization and gene expression in the cortex of Neurod2 null mice. | Molecular Psychiatry

Fig. 1: Altered laminar organization and gene expression in the cortex of Neurod2 null mice.

From: Disruption of NEUROD2 causes a neurodevelopmental syndrome with autistic features via cell-autonomous defects in forebrain glutamatergic neurons

Fig. 1

a–i Altered lamination of cortical layers at P30 in Neurod2 KO mice. a Neurod2 mRNA and NEUROD2 protein were expressed from corticogenesis to adulthood. In situ hybridization pictures are from ALLEN BRAIN ATLAS. CP: cortical plate, IZ: intermediate zone, MZ: marginal zone, V/SVZ: ventricular/subventricular zone. b Representative confocal photomicrographs of S1 cortical columns at the somatosensory level for the four markers analyzed. c–f Left, mean cell laminar position in percentage of the cortical thickness. Right, twenty-bin based laminar distribution of cPN subtypes in WT (black bars) and KO mice (red bars). We used the markers TBR1 for L6 (c), BCL11B for L5 (d), RORB for L4 (e) and CUX1 for L2-4 (f). We analyzed 25 to 42 S1 cortical columns from 9/6/9/7 WT and 5/5/5/5 KO mice (for TBR1, BCL11B, RORB and CUX1, respectively). g Probability density function showing the average laminar distribution of cells. All layers were switched superficially, with the strongest significance for L5 and L4. h–k Dysregulation of layer-specific genes in Neurod2 KO mice. (h) Number of cPN subtypes in a S1 cortical column, labeled using the four-layer markers; note the absence of significant difference between genotypes. i Thickness of the BCL11B, RORB, and CUX1 layers. j–m Differential expression in Neurod2 KO S1-M1 cortex at P30. j Schematic of the RNA-seq experiment. k Volcano plots depicting differential gene expression. Right plot is a magnification of left plot (N = 3 biological replicates). l Venn diagram of DEX genes showing enrichement in L2/3, L5, and L6 (180 are generic to all layers, some are common between 2 layers and others specific to a single layer). m Fold change expression (FC; log2 scale) of DEX genes showing layer enrichment in normal conditions. Left bars depict downregulated genes, right bars show upregulated genes. Genes are ranked according to adjusted P value). Data are represented as means ± SEM except in mean cell position plots (c–f) where median, first and third quartiles are represented. Statistical significance was evaluated by permutation test for medians [(c–f), (h–i)], Student t-test or Mann–Whitney test depending on normality of samples [(h–i)], Anderson–Darling test [probability densities in (g)] or by Permutation test for a spatially adjusted two-way ANOVA followed by Bonferroni’s post hoc test [bin graphs in (c–f)] (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). See also Figs. S1–S3.

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