Generation of an enteric smooth muscle cell line from pig ileum
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA621635 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11458987_1.fastq.gz | Fastqsanger.gz | |||
| SRR11458987_2.fastq.gz | Fastqsanger.gz | |||
| SRR11458988_1.fastq.gz | Fastqsanger.gz | |||
| SRR11458988_2.fastq.gz | Fastqsanger.gz |
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Project description:The goal of this study was to characterize a single-cell clone derived from pig ileum epithelium. Analyses including RNA-seq demonstrated that this clone was derived from an enteric smooth muscle cell. This clone is named PIC7 in the manuscript.
2020-04-29 | GSE147822 | GEO
Project description:Smooth muscle cells (SMCs) play an important role in physiology and production in farm animals such as pigs. Here, we report the generation of a pig SMC line. Our original objective was to establish an enteroendocrine cell line from the pig ileum epithelium through lentiviral transduction of the Simian Virus (SV) 40 large T antigen. However, an initial expression analysis of marker genes in nine cell clones revealed that none of them were enteroendocrine cells or absorptive enterocytes, goblet cells, or Paneth cells, some of the major cell types existing in the ileum epithelium. A more detailed characterization of one clone named PIC7 by RNA-seq showed that these cells expressed many of the known smooth muscle-specific or -enriched genes, including smooth muscle actin alpha 2, calponin 1, calponin 3, myosin heavy chain 11, myosin light chain kinase, smoothelin, tenascin C, transgelin, tropomyosin 1, and tropomyosin 2. Both quantitative PCR and RNA-seq analyses showed that the PIC7 cells had a high expression of mRNA for smooth muscle actin gamma 2, also known as enteric smooth muscle actin. A Western blot analysis confirmed the expression of SV40 T antigen in the PIC7 cells. An immunohistochemical analysis demonstrated the expression of smooth muscle actin alpha 2 filaments in the PIC7 cells. A collagen gel contraction assay showed that the PIC7 cells were capable of both spontaneous contraction and contraction in response to serotonin stimulation. We conclude that the PIC7 cells are derived from an enteric SMC from the pig ileum. These cells may be a useful model for studying the cellular and molecular physiology of pig enteric SMCs. Because pigs are similar to humans in anatomy and physiology, the PIC7 cells may be also used as a model for human intestinal SMCs.
| S-EPMC7179811 | biostudies-literature
Project description:We used the ileal loop model to assess the effects of enteric bacteria organisms on host gene expression in intestinal tissue independent of and following early SIV infection. SIV infection in the gut causes rapid and severe immune dysfunction and damage to the intestinal structure, this may alter the intimate interaction with lumenal organisms. This study was performed to determine whether early SIV infection, prior to the depletion of CD4+ T cells, can alter interaction of the host with pathogenic Salmonella serovar Typhimurium (ST) or commensal Lactobacillus plantarum (LP), and to further understand the earliest changes to the intestinal mucosa following SIV infection. We used microarray analysis to detail the global program of gene expression underlying changes in the ileum following early SIV infection, and if these changes in any way alter the host interaction/ response to pathogenic and commensal enteric bacteria. A subset of animals were infected with SIVmac251 for 2.5 days, following which they underwent a loop surgery where the ileum was sectioned off with surgical ties and bacteria (ST or LP) or their respective media controls (LB or MRS) were injected intralumenally. Following 5 hours of incubation the loops were excised and frozen. Tissue sections were cut and RNA extracted for gene expression analysis. Rhesus Macaques infected with SIVmac251 for 2.5 days compared to uninfected controls. Comparisons are of ileum loop sections injected with Salmonella serovar Typhimurium, Lactobacillus Plantarum or respective media controls LB or MRS.
2014-12-19 | E-GEOD-52589 | biostudies-arrayexpress
Project description:We used the ileal loop model to assess the effects of enteric bacteria organisms on host gene expression in intestinal tissue independent of and following early SIV infection. SIV infection in the gut causes rapid and severe immune dysfunction and damage to the intestinal structure, this may alter the intimate interaction with lumenal organisms. This study was performed to determine whether early SIV infection, prior to the depletion of CD4+ T cells, can alter interaction of the host with pathogenic Salmonella serovar Typhimurium (ST) or commensal Lactobacillus plantarum (LP), and to further understand the earliest changes to the intestinal mucosa following SIV infection. We used microarray analysis to detail the global program of gene expression underlying changes in the ileum following early SIV infection, and if these changes in any way alter the host interaction/ response to pathogenic and commensal enteric bacteria.
2014-12-19 | GSE52589 | GEO
Project description:To identify the gene expression profile of enteric glia and assess the transcriptional similarity between enteric and extraenteric glia, we performed RNA sequencing analysis on PLP1-expressing cells in the mouse intestine. This analysis shows that enteric glia are transcriptionally unique and distinct from other cell types in the nervous system. Enteric glia express many genes characteristic of the myelinating glia, Schwann cells and oli- godendrocytes, although there is no evidence of myelination in the murine ENS. Total RNA expression profiles of PLP1 expressing enteric glial cells (GFP+) and non-glial cells (GFP-negative) were obtained from the ileum and colon of juvenile PLP1-eGFP transgenic mice.
2015-09-01 | E-GEOD-72560 | biostudies-arrayexpress
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
Project description:Intrinsic enteric neurons (iENs) form a crucial neuronal network within the myenteric and submucosal plexus of the gastrointestinal tract, primarily responsible for regulating gut peristalsis. The mechanisms by which iENs sense and integrate dietary and microbial signals to regulate intestinal homeostasis and inflammation remain unclear. Here we showed that environmental sensor aryl hydrocarbon receptor (AHR) was expressed in different iEN subsets in the ileum and colon, and AHR ligands differentially modulated iEN activity in these regions. Genetic perturbation of Ahr in neurons increased iEN activation in the ileum but conversely decreased it in the colon in response to different intestinal pathogens. Furthermore, neuronal AHR deficiency enhanced the clearance of bacterial pathogens, which was associated with increased proliferation and abundance of group 3 innate lymphoid cells in the ileum. Together, our findings demonstrate the region-specific functions of AHR in neurons in response to infections.
2025-08-19 | GSE290795 | GEO
Project description:To use surgical resection specimens of the intact bowel wall from inflammatory bowel disease (IBD) patients and perform RNA-seq, bioinformatics analysis, and validation by quantitative polymerase chain reaction (qPCR) and immunohistochemistry (IHC) to explore the subtle differences between Crohn's disease (CD) and ulcerative colitis (UC), with predominant focus on the smooth muscle cells (SMCs) and enteric autonomic nervous system (ANS).
2023-03-28 | GSE227747 | GEO
Project description:Microbial density and diversity increase in distal intestinal segments, affecting tissue physiology, metabolism, and function of both the immune and nervous systems. We characterized the influence of the microbiota on murine intrinsic enteric-associated neurons (iEAN). We found that iEAN are functionally adapted to the intestinal segment they occupy, with a stronger microbiota influence on ileal and colonic neurons. Chemogenetic characterization of microbiota-influenced iEAN identified a subset of viscerofugal CART+ neurons, enriched in the ileum and colon, able to modulate feeding and glucose metabolism. Retro- and anterograde tracing revealed that CART+ viscerofugal neurons send axons to the prevertebral ganglia and are poly-synaptically connected to the liver and pancreas. Microbiota depletion led to NLRP6 and Caspase 11-dependent loss of CART+ neurons, and impaired liver-mediated gluconeogenesis. Our results demonstrate a region-specific adaptation of enteric neurons and indicate that iEAN subsets are capable of regulating blood glucose levels independently from the central nervous system.
2020-08-13 | GSE156142 | GEO
Project description:Deoxynivalenol (DON) is one of the frequent Fusarium mycotoxins and poses a serious threat to public health worldwide, especially in humans and pigs. Growth retardation is the critical symptom of DON poisoning, commonly manifested by weight loss and reduced food intake. The gastrointestinal tract (GIT) is the principal target organ in the body and DON-induced weight loss is tightly connected with its capacity to reduce feed intake by disturbing the GIT motility. Our previous reports for the first time indicated that DON interfered with intestinal motility by injuring the contractility of enteric smooth muscle cells (SMC), suggesting that enteric SMC might be a main target in the procession of DON-induced GIT motility disorder. However, underlying molecular mechanisms of DON-induced enteric SMC contractile dysfunction remain poorly understood until now. Here, we explored the potential molecular mechanisms by employing a complementary method of transcriptomics and proteomics using the porcine enteric smooth muscle cell line (PISMC) as an experimental model. First, the DON-induced contractile dysfunction of the enteric SMC model was successfully established with PISMC, which was evidenced by decreased cell contractility, cell viability, and the expression levels of SMC contractile markers (contractile apparatus components). Subsequently, transcriptomic and proteomic analyses were used to reveal the differentially expressed gene and protein profiles after DON exposure. A total of 2944 genes and 980 proteins were identified as differentially expressed. Additionally, a systematic bioinformatic analysis of gene-protein interactions revealed that a total of 152 co-differentially expressed genes were identified. The data uncover that the expression of numerous extracellular matrix (ECM) proteins was downregulated in PISMC under DON exposure. DON also remarkably decreased the expression of multiple integrin subunits, resulting in the inhibition of the ECM-integrin receptor interaction and its mediated signaling. Furthermore, DON treatment could depress actin polymerization, as reflected by the upregulated expression of Rho GTPase-activating proteins, cofilin and the downregulated expression levels of Rho guanine nucleotide exchange factors in PISMC. Meanwhile, the expression levels of downstream target genes that were responsible for sustaining contractile phenotype were significantly inhibited in PISMC challenge with DON, including actin alpha 2, caldesmon 1, and myosin regulatory light chain 9 and so on. Taken together, the current results suggest that DON inhibits enteric smooth muscle cell contractility via regulating the ECM-integrin-actin polymerization signaling pathway mediated downstream contractile apparatus genes expression. Our findings provide novel insights into potential mechanisms behind the DON toxicological effects in the GIT of humans and animals.
2023-03-06 | PXD040605 |