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ABSTRACT: RNASeq analysis of embryonic day 13.5 mouse cerebral cortex from (i) Pax6-deleted and (ii) Pax6- and Gsx2-deleted embryos

PROVIDER: PRJEB21105 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1995120_1.fastq.gz	Fastqsanger.gz
	ERR1995120_2.fastq.gz	Fastqsanger.gz
	ERR1995130_1.fastq.gz	Fastqsanger.gz
	ERR1995130_2.fastq.gz	Fastqsanger.gz
	ERR1995121_1.fastq.gz	Fastqsanger.gz

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Project description:RNASeq analysis of embryonic day 13.5 mouse cerebral cortex from control and Pax6-deleted embryos

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Project description:Pax6 is a transcription factor with key functional roles in embryonic development. In order to identify downstream effectors of Pax6 in the developing cerebral cortex we performed microarray analysis. We compared gene expression profiles of cortical tissues isolated from wild type and Pax6-/- mouse embryos. In order to identify Pax6 downstream targets we carried out microarray analysis of Pax6-/- mutant mice. Pax6 is highly expressed in the mouse cerebral cortex at embryonic day E14.5, therefore we selected this tissue in order to compare gene expression profiles between wild type and Pax6-/- homozygous cortici. RNA samples were isolated from three mutant and three wild type embryos.

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Evidence that Dmrta2 acts through repression of Pax6 in cortical patterning and identification of a mutation crucial for DNA recognition associated with microcephaly in human

Project description:Dmrta2 (also designated Dmrt5) is a transcriptional regulator expressed in cortical progenitors in a caudomedial high/rostrolateral low gradient with important roles at different steps of cortical development. Dmrta2 has been suggested to act in cortex development mainly by differential suppression of Pax6 and other homeobox transcription factors such as the ventral telencephalic regulator Gsx2, which remains to be fully demonstrated. Here we have addressed the epistatic relation between Pax6 and Dmrta2 by comparing phenotypes in mutant embryos or embryos overexpressing both genes in various allelic combinations. We show that Dmrta2 cooperates with Pax6 in the maintenance of cortical identity in dorsal telencephalic progenitors and that it acts as a transcriptional repressor of Pax6 to control cortical patterning. Mechanistically, we show that in P19 cells, Dmrta2 acts as a DNA14 binding dependent repressor on the Pax6 E60 enhancer and that a point mutation that affects its DNA binding properties identified in a consanguineous family leads to agenesis of the corpus callosum, pachygyria, and the absence of the cingulate gyrus. Finally, we provide evidence that Dmrta2 binds components of the NuRD repressor complex and interacts with zinc finger proteins such as Zfp423. Together, our results highlight the importance and conserved function of Dmrta2 in cortical development and provide novel insights into its mechanism of action.

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NeuroD2 ChIP-SEQ from embryonic cortex

Project description:NeuroD2 targets were identified from embryonic day 14.5 cerebral cortex tissue. The cerebral cortex (dorsal telencelphalon) from E14.5 mouse embryos was dissected and ChIP-SEQ was performed using three separate antibodies against NeuroD2.

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Project description:To investigate the differences in gene expression among various cells and tissues, hippocampus, cerebellum, cortex, primary glial cells and primary neurons were isolated from mice, mouse embryonic fibroblasts (MEFs) were isolated from day 13.5 embryos, R1 mbryonic stem cells (ESCs) were clutured in galetin-coated plates in N2B27+2i medium. Total RNA were extracted and subjected to RNA-Seq analysis.

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Project description:High histone acetylation is associated with high transcriptional activity. The lysine acetyltransferase KAT6B is known to be required for histone acetylation and KAT6B is essential for normal brain development. In this study we examined the effects of loss and gain of KAT6B on gene expression in the developing cerebral cortex. We isolated RNA from the dorsal telencephalon of embryonic day 12.5 embryos, which is the primordium of the cerebral cortex, and from the E15.5 foetal cortex of mouse embryos and foetuses that lacked KAT6B or overexpressed KAT6B. Genes required for brain development and neuronal differentiation were downregulated in Kat6b null tissues and upregulated in Kat6b transgenic overexpressing tissue.

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