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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Similar Datasets
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:Muscarinic-receptor stimulation by 0.1 mM-carbachol in longitudinal muscle of the guinea-pig ileum increases the incorporation of [3H]inositol into inositol-containing phospholipid. This effect was blocked by 16 microM-atropine. After 60 min incubation, carbachol increased the accumulation of total inositol phosphates 20-fold in the presence of 10 mM-Li+. Less than 20% of the total inositol phosphate corresponded to inositol 1-phosphate by ion-exchange chromatography, whereas of the remainder about two-thirds corresponded to inositol bisphosphate and one third to inositol trisphosphate. It is concluded that stimulation of muscarinic receptors in guinea-pig ileum enhances breakdown of polyphosphoinositides, suggesting that this may be a primary event associated with Ca2+ mobilization in the guinea-pig ileum.
| S-EPMC1144327 | biostudies-other
Project description:Thiele2013 - Smooth muscle smooth muscle cells
The model of smooth muscle smooth muscle cells metabolism is derived from the community-driven global reconstruction of human metabolism (version 2.02, MODEL1109130000
).
This model is described in the article:
A community-driven global reconstruction of human metabolism.
Thiele I, et al
.
Nature Biotechnology
Abstract:
Multiple models of human metabolism have been reconstructed, but each represents only a subset of our knowledge. Here we describe Recon 2, a community-driven,
consensus 'metabolic reconstruction', which is the most comprehensive representation of human metabolism that is applicable to computational modeling. Compared
with its predecessors, the reconstruction has improved topological and functional features, including ~2x more reactions and ~1.7x more unique metabolites. Using
Recon 2 we predicted changes in metabolite biomarkers for 49 inborn errors of metabolism with 77% accuracy when compared to experimental data. Mapping metabolomic
data and drug information onto Recon 2 demonstrates its potential for integrating and analyzing diverse data types. Using protein expression data, we automatically
generated a compendium of 65 cell type-specific models, providing a basis for manual curation or investigation of cell-specific metabolic properties. Recon 2 will
facilitate many future biomedical studies and is freely available at http://humanmetabolism.org/.
This model is hosted on BioModels Database
and identified by: MODEL1310110025
.
To cite BioModels Database, please use: BioModels Database: An enhanced,
curated and annotated resource for published quantitative kinetic models
.
To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer
to CC0 Public Domain Dedication
for more information.
2005-01-01 | MODEL1310110025 | BioModels
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: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 |