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Cell-autonomous retinoic acid receptor signaling alters enteric nervous system development with stage-specific effects in mice

ABSTRACT: Cell-autonomous retinoic acid receptor signaling alters enteric nervous system development with stage-specific effects in mice

PROVIDER: PRJNA694135 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13509460.fastq.gz	Fastqsanger.gz
	SRR13509461.fastq.gz	Fastqsanger.gz
	SRR13509462.fastq.gz	Fastqsanger.gz
	SRR13509463.fastq.gz	Fastqsanger.gz
	SRR13509464.fastq.gz	Fastqsanger.gz

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Cell-autonomous retinoic acid receptor signaling alters enteric nervous system development with stage-specific effects in mice

Project description:Purpose: Determine how blocking retinoic acid receptor (RAR) signaling within enteric neural crest-derived cells (ENCDC) from E11.5 stomach or E13.5 colon alters mRNA abundance. These ENCDC become the enteric nervous system. Methods: Enteric nervous system precursors were isolated from mouse E11.5 stomach or E13.5 colon using fluorescence activated cell sorting to purify TdTomato or EYFP expressing cells respectively from fetal bowel. The RarαDN allele encodes a potent RAR dominant negative protein that is expressed after CRE-mediated DNA recombination. Tamoxifen-activated CRE-ERT2 was expressed from the Ret locus for E13.5 colon studies and mice were tamoxifen treated at E10.5 prior to cell sorting. The Wnt1Cre allele was employed for E11.5 stomach studies. RNA-SEQ was performed in quadruplicate using Illumina HiSeq 4000. Sequence reads that passed quality filters were aligned to remove repeat sequences and ribosomal RNA reads and then processed using RNA-Seq unified mapper (RUM) package. RUM files were visualized in the TessLA browser for subsequent analyses. Results: Transcriptional profiling shows RarαDN differentially impacts gene expression in E11.5 stomach and E13.5 colon enteric neural crest-derived cells (ENCDC) that become the enteric nervous system. Conclusions: Blocking of RAR signaling in ENCDC causes dramatic changes in gene expression during early development in a stage-specific manner.

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Cell-autonomous retinoic acid receptor signaling has stage-specific effects on mouse enteric nervous system.

Project description:Retinoic acid (RA) signaling is essential for enteric nervous system (ENS) development, since vitamin A deficiency or mutations in RA signaling profoundly reduce bowel colonization by ENS precursors. These RA effects could occur because of RA activity within the ENS lineage or via RA activity in other cell types. To define cell-autonomous roles for retinoid signaling within the ENS lineage at distinct developmental time points, we activated a potent floxed dominant-negative RA receptor α (RarαDN) in the ENS using diverse CRE recombinase-expressing mouse lines. This strategy enabled us to block RA signaling at premigratory, migratory, and postmigratory stages for ENS precursors. We found that cell-autonomous loss of RA receptor (RAR) signaling dramatically affected ENS development. CRE activation of RarαDN expression at premigratory or migratory stages caused severe intestinal aganglionosis, but at later stages, RarαDN induced a broad range of phenotypes including hypoganglionosis, submucosal plexus loss, and abnormal neural differentiation. RNA sequencing highlighted distinct RA-regulated gene sets at different developmental stages. These studies show complicated context-dependent RA-mediated regulation of ENS development.

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Project description:Expression analysis of adult zebrafish enteric nervous system

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Enteric glia express proteolipid protein 1 and are a transcriptionally unique population of glia in the mammalian nervous system

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