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ScRNA-sequencing reveals new enteric nervous system roles for GDNF, NRTN, and TBX3

ABSTRACT: scRNA-sequencing reveals new enteric nervous system roles for GDNF, NRTN, and TBX3

PROVIDER: PRJNA659493 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12520100.fastq.gz	Fastqsanger.gz
	SRR12520101.fastq.gz	Fastqsanger.gz
	SRR12520102.fastq.gz	Fastqsanger.gz
	SRR12520103.fastq.gz	Fastqsanger.gz
	SRR12520104.fastq.gz	Fastqsanger.gz

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scRNA-sequencing reveals new enteric nervous system roles for GDNF, NRTN, and TBX3

Project description:Bowel function requires coordinated activity of many enteric neuron subtypes. Clear definition of subtype-specific gene expression may facilitate molecular diagnoses for bowel motility disorders. Using adult mouse colon RNAseq data from 635 myenteric neurons and 707 E17.5 neurons, we defined seven adult myenteric neuron subtypes, eight E17.5 neuron subtypes and hundreds of differentially-expressed genes. Manually dissected human colon myenteric plexus yielded data from 48 neurons, 3798 glia, 5568 smooth muscle, 377 interstitial cells, and 2153 macrophages. Immunohistochemistry demonstrated differential protein abundance for BNC2, PBX3, RBFOX1, TBX2, and TBX3 in enteric neuron subtypes. Conditional Tbx3 loss reduced NOS1-expressing myenteric neurons. Differential Gfra1 and Gfra2 expression coupled with calcium imaging revealed that GDNF and neurturin acutely and differentially regulate activity of ~50% of myenteric neurons with distinct effects on smooth muscle contractions. This insight into enteric nervous system biology provides a foundation for future studies of bowel motility disorders.

2021-01-01 | GSE156905 | GEO

scRNA-Seq Reveals New Enteric Nervous System Roles for GDNF, NRTN, and TBX3.

Project description:Background and aimsBowel function requires coordinated activity of diverse enteric neuron subtypes. Our aim was to define gene expression in these neuron subtypes to facilitate development of novel therapeutic approaches to treat devastating enteric neuropathies, and to learn more about enteric nervous system function.MethodsTo identify subtype-specific genes, we performed single-nucleus RNA-seq on adult mouse and human colon myenteric plexus, and single-cell RNA-seq on E17.5 mouse ENS cells from whole bowel. We used immunohistochemistry, select mutant mice, and calcium imaging to validate and extend results.ResultsRNA-seq on 635 adult mouse colon myenteric neurons and 707 E17.5 neurons from whole bowel defined seven adult neuron subtypes, eight E17.5 neuron subtypes and hundreds of differentially expressed genes. Manually dissected human colon myenteric plexus yielded RNA-seq data from 48 neurons, 3798 glia, 5568 smooth muscle, 377 interstitial cells of Cajal, and 2153 macrophages. Immunohistochemistry demonstrated differential expression for BNC2, PBX3, SATB1, RBFOX1, TBX2, and TBX3 in enteric neuron subtypes. Conditional Tbx3 loss reduced NOS1-expressing myenteric neurons. Differential Gfra1 and Gfra2 expression coupled with calcium imaging revealed that GDNF and neurturin acutely and differentially regulate activity of ∼50% of myenteric neurons with distinct effects on smooth muscle contractions.ConclusionSingle cell analyses defined genes differentially expressed in myenteric neuron subtypes and new roles for TBX3, GDNF and NRTN. These data facilitate molecular diagnostic studies and novel therapeutics for bowel motility disorders.

| S-EPMC8099699 | biostudies-literature

RNA Expression Data from developing enteric nervous system (ENS) and bowel

Project description:The enteric nervous system (ENS) can control most essential gut functions owing to its organization into complete neural circuits consisting of a multitude of different neuronal subtypes. We used microarrays to identify transcription factor networks and signaling pathways involved in diversification and differentiation of enteric neurons during development of the enteric nervous system.

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Expression analysis of adult zebrafish enteric nervous system

Project description:Expression analysis of adult zebrafish enteric nervous system

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Expression analysis of adult zebrafish enteric nervous system

Project description:The enteric nervous system (ENS) encompasses the intrinsic neuroglia networks of the gastrointestinal (GI) tract that are essential for digestive function and gut homeostasis. To investigate the ENS of zebrafish, we carried out bulk RNA sequencing on nuclei purified by FACS (fluorescent-activated cell sorting) representing both the Cherry+ (ENS) and Cherry- (non-ENS) muscularis externa cell populations of Tg(sox10:Cre;Cherry) zebrafish gut.

2020-09-14 | GSE145885 | GEO

Profiling chromatin accessibility in Enteric Nervous System progenitors during neurogenic phases

Project description:We utilized a Phox2b-H2BCerulean transgene that is expressed at low levels in Enteric Nervous System progenitors (ENPs) and enteric glia that is also expressed at notably higher levels in differentiating enteric neurons to capture these populations during neurogenic phases of ENS development. Collected tissues from 16.5 days post coitus developing mouse intestine included the stomach, small intestine, and colon that were dissociated to single cell suspensions for flow sort capture of Phox2b-H2BCerulaen+ cells. We applied differential gating in flow sorts to capture populations expressing low levels of the Phox2b-H2BCerulaen transgene (ENPs and enteric glia) and while concurrently collecting cells expressing high levels of the Phox2b-H2BCerulaen transgene (developing and maturing enteric neurons) from the same samples. Nuclei were generated from these Phox2b-H2BCerulaen “high” and “low” populations and encapsulated separately to produce single nucleus ATAC-Seq libraries using 10X Genomics chemistry. Sequencing was performed on the Illumina NovaSeq6000 (S4) using PE150 Sequencing targeting >50,000 reads per nucleus. Comparison of the resulting sequence data from the “high” versus “low” cell populations allowed us to identify differentially accessible genome regions in developing enteric neurons compared to ENPs and enteric glia.

2026-02-05 | GSE298406 | GEO

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