Project description:To date, speculations on the molecular roles of Tbr2 transcription factor during specification, maintenance and differentiation of cortical specific Intermediate Neural Progenitors are coming from the analysis of loss- and gain-of-function experiments. However since its capacity to bind the DNA to extert its function we did Chromatin Immuno-Precipitation followed by deep sequencing to profile its target on the genome. We performed this approach directly in vivo to respect the physiological and peculiar enviroment of its action during cortical development. Moreover we did the same approach for Neurogenin 2 proneural gene. Interestingly the vast majority of the Neurog2 peaks are in common with Tbr2 dataset suggesting a co-operation between the two factors confirmed by biochemical and functional assays. Finally we compared Tbr2 dataset with the DNA regions bound by the H3K27me3 histone demethylase Jmjd3 (NCBI:Kdm6b) obtained by others. Interestingly we were able to find regions in common linked to important genes for neuronal differentiation. 3 samples, one input chromatin, one ChIP for Tbr2 and one ChIP for Neurog2
Project description:To date, speculations on the molecular roles of Tbr2 transcription factor during specification, maintenance and differentiation of cortical specific Intermediate Neural Progenitors are coming from the analysis of loss- and gain-of-function experiments. However since its capacity to bind the DNA to extert its function we did Chromatin Immuno-Precipitation followed by deep sequencing to profile its target on the genome. We performed this approach directly in vivo to respect the physiological and peculiar enviroment of its action during cortical development. Moreover we did the same approach for Neurogenin 2 proneural gene. Interestingly the vast majority of the Neurog2 peaks are in common with Tbr2 dataset suggesting a co-operation between the two factors confirmed by biochemical and functional assays. Finally we compared Tbr2 dataset with the DNA regions bound by the H3K27me3 histone demethylase Jmjd3 (NCBI:Kdm6b) obtained by others. Interestingly we were able to find regions in common linked to important genes for neuronal differentiation.
Project description:The abscence of TBR2 gene in human leads to microcephaly. This condition is mimicked by the specific ablation of the murine gene in developing cerebral cortex. Herein we compared gene expression in control and Tbr2 cKO in E14.5 cerebral cortices. This approach represents a useful tool to identify the molecular mechanisms at the basis of the phenotype. 6 samples, 3x Tbr2 +/+;Foxg1::Cre (control) and 3x Tbr2 fl/fl;Foxg1::Cre
Project description:The abscence of TBR2 gene in human leads to microcephaly. This condition is mimicked by the specific ablation of the murine gene in developing cerebral cortex. Herein we compared gene expression in control and Tbr2 cKO in E14.5 cerebral cortices. This approach represents a useful tool to identify the molecular mechanisms at the basis of the phenotype.
Project description:To identify and purify NEUROG2-expressing cells and trace their short-term lineage, we engineered two NEUROG2-mCherry knock-in human embryonic stem cell (hESC) lines. Transcriptomic profiling of NEUROG2:mCherry knock-in hESC-derived cerebral organoids revealed an enrichment of neurogenic, oligodendrocyte precursor cell and extracellular matrix-associated gene transcripts in mCherry-high cells. Conversely, only neurogenic gene transcripts were enriched in mCherry-high cells from Neurog2:mCherry knock-in mouse cortices.
Project description:Humans exemplify gyrencephalic species with folded cerebral cortices, contrasting with lissencephalic mammals such as mice. Here we investigated how proneural genes Neurog2 and Ascl1 control cortical folding by regulating neurogenic patterns. Cortical neural progenitor cells (NPCs) stratify into four pools (proneural negative, Neurog2+, Ascl1+, double+) that are distributed evenly in mouse cortices and modular in gyrencephalic macaque cortices and pseudo-folded human cerebral organoids. Each pool has distinct developmental potentials, transcriptomes, epigenomes, and gene regulatory networks. Neurog2-Ascl1 form a bistable toggle switch double+ NPCs to prevent lineage commitment observed in single+ NPCs. Neurog2 and Ascl1 act redundantly to control neurogenic timing, with NPCs precociously depleted in Neurog2-/-;Ascl1-/- cortices. Finally, selective killing of Neurog2/Ascl1 double+ NPCs using Neurog2/Ascl1 split-Cre;Rosa-DTA transgenics breaks neurogenic symmetry in mice by locally disrupting Notch signaling, leading to cortical folding. Our findings suggest that Neurog2/Ascl1 double+ NPCs are Notch-ligand expressing ‘niche’ cells that regulate neurogenic continuity and cortical gyrification.
Project description:Taf8 was deleted in the mouse central nervous system using our Taf8 conditional allele. To determine transcriptional changes due to Ta8 deletion in the developing CNS, unstranded RNA sequencing was performed on cortices from Taf8 deleted embryos (E14.5 Taf8lox/–NesCreT/+p53–/–) and control embryos (E14.5 Taf8+/+NesCreT/+p53–/–).