DNA methylation analysis of porcine mammary epithelial cells challenged with Escherichia coli
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Illumina HiSeq 2500
ORGANISM(S): Sus scrofa
SUBMITTER: Nares Trakooljul
PROVIDER: E-MTAB-7363 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Porcine mammary epithelial cell (PMEC) cultures of three lactating sows were treated with potential mastitis-causing pathogens E. coli and S. aureus in vitro. Subsequently transcriptome profiles were analysed after 3 h and 24 h post-challenge, respectively. per experimental group 3 PMEC cultures were sampled in triplicate for expression profiling.
2015-12-18 | E-GEOD-64246 | biostudies-arrayexpress
Project description:Obtaining an in depth understanding of the arms races between peptides comprising the innate immune response and bacterial pathogens is of fundamental interest and will inform the development of new antibacterial therapeutics. Many cationic antimicrobial peptides (AMPs) share a range of structural and physical features that have been linked to antibacterial activity and yet they vary dramatically in their potency towards the same bacterial target. We hypothesised that a whole organism view of AMP challenge on Escherichia coli could provide a sophisticated, bacterial perspective enabling understanding of how potency is linked to mode of action. We used a 1H NMR metabolomic approach to characterise the effect on E. coli of challenge with four structurally and physically related AMPs: magainin 2, pleurocidin, buforin II and a designed peptide comprising D-amino acids only. Sub-inhibitory conditions, where these peptides nevertheless induced a bacterial response, were identified enabling electron microscopic and transcriptomic analyses. Although some common features of the bacterial response to AMP challenge could be identified, the metabolomes, morphological changes and the vast majority of the changes in gene expression were specific to each AMP. We show the antibacterial mode of action of AMPs can be accurately predicted by comparing ontological profiles generated by transcriptomic analyses. The response of E. coli to AMP challenge is highly plastic, with the bacteria capable of deploying a multifaceted response adapted to the mode of action rather than the physical properties of the AMP.
2014-01-21 | E-MTAB-1703 | biostudies-arrayexpress
Project description:We performed a genome-wide transcriptional analysis in the mammary gland in a mouse model of E. coli mastitis using high-density mouse oligonucleotide microarrays. This global transcription analysis revealed that about 7% of tested genes are mobilized in the mouse mammary gland to E. coli endotoxin. We identified 1402 differentially expressed genes that were associated with physiological system development/function and molecular/cellular functions and metabolic/signalling pathways that are highly relevant to host immune-inflammatory defense response against E. coli infection. The mouse differentially expressed genes through the use of comparative mapping/genomics and positional information on reported QTL for bovine mastitis allowed identifying 293 potential candidate genes for bovine mastitis. This study will enable other researchers to combine our mRNA expression data with genetic association studies to discover genomic variation underlying variation of susceptibility to mastitis in dairy cows. Keywords: time course, disease state analysis This study was conducted using a split-plot design. We measured levels of gene expression in healthy (saline injected) and diseased (E. coli injected) mammary gland tissue (treatment = 2, in sub-plots). The mammary tissue samples were collected at two-time points: 24 hr and 48 hr post-injection (time = 2, in main-plots). Each inbred mouse was injected with both treatments: the 4th left mammary gland was injected with saline solution and the 4th right mammary gland was injected with E. coli solution (mice = 4). The expression profiling was carried out using two biological replications (replication = 2) at each time point.
2011-06-29 | E-GEOD-14485 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series:; GSE15019: Transcriptome analysis of bovine mammary gland tissue treated with E. coli for 6 hours; GSE15020: Transcriptome analysis of bovine mammary gland tissue treated with E. coli for 24 hours; GSE15022: Transcriptome analysis of bovine mammary gland tissue of an udder quarter adjacent to an E.coli treated one for 24 hrs Experiment Overall Design: Refer to individual Series
2009-06-08 | E-GEOD-15025 | biostudies-arrayexpress
Project description:By comparison of the transcriptome profiles of infected and healthy udder tissue we analyse gene expression in the early stage of infection with E. coli 1303. Differentially expressed genes and transcription factors regulating coexpressed genes identified by SOTA clustering were used to identify key genes that define early stage of infection with E. coli 1303. Experiment Overall Design: 10 samples, two conditions: E. coli treated for 6h vs healthy control * five replicates
2009-06-08 | E-GEOD-15019 | biostudies-arrayexpress
Project description:By comparison of the transcriptome profiles of infected and healthy udder tissue we analyse gene expression in the late stage of infection with E. coli 1303. Differentially expressed genes and transcription factors regulating coexpressed genes identified by SOTA clustering were used to identify key genes that define late stage of infection with E. coli 1303. Experiment Overall Design: 10 samples, two conditions: E. coli treated for 24h vs healthy control * five replicates
2009-06-08 | E-GEOD-15020 | biostudies-arrayexpress
Project description:By comparison of the transcriptome profiles of udder quarters neighboring to infected quarters and healthy udder tissue we analyse gene expression in the late stage of infection with E. coli 1303. Differentially expressed genes and transcription factors regulating coexpressed genes identified by SOTA clustering were used to identify key genes that define systemic reactions to infection with E. coli 1303. Experiment Overall Design: 10 samples, two conditions: udder quarters neighboring to one which was treated with E. coli for 24h vs healthy control * five replicates
2009-06-08 | E-GEOD-15022 | biostudies-arrayexpress
Project description:Measuring precise concentrations of proteins can provide insights into biological processes. Here we use efficient protein extraction and sample fractionation, as well as state-of-the-art quantitative mass spectrometry techniques to generate a comprehensive, condition-dependent protein-abundance map for Escherichia coli. We measure cellular protein concentrations for 55% of predicted E. coli genes (>2,300 proteins) under 22 different experimental conditions and identify methylation and N-terminal protein acetylations previously not known to be prevalent in bacteria. We uncover system-wide proteome allocation, expression regulation and post-translational adaptations. These data provide a valuable resource for the systems biology and broader E. coli research communities.
2015-10-12 | PXD000498 | Pride
Project description:Adult pituitary stem cells (SC) are mainly dormant, yet in the case of damage the resident SC compartment shows swift activation. More in particular, following diphtheria toxin (DT)-triggered endocrine cell-ablation injury in the GHCre/iDTR mouse model (expressing the Cre recombinase under control of the Gh promoter, as well as the Cre-inducible DT receptor (iDTR)), the pituitary stem cell population is promptly activated displaying enhanced proliferative activity and expansion. Substantial regeneration of the depleted somatotropes is eventually observed after 5 to 6 months. However, this regenerative competence rapidly drops with age, and is no longer observed when mice get older (from 8-10 months of age). Virtually nothing is known about the molecular machinery driving the quiescent pituitary stem cells into activation (as observed following local injury), neither about how this route may change at advancing age. Here, we started to tackle this query by interrogating young (9-11 weeks) and old (14 months), damaged and undamaged pituitary (specifically, its major endocrine AP lobe) using single-cell RNA sequencing. We focused on the prompt stem cell reaction that occurs immediately upon the DT-induced local damage in the GHCre/iDTR model.
2021-06-15 | E-MTAB-10021 | biostudies-arrayexpress
Project description:GCC was used to determine the structure of E. coli grown in LB or treated with SHX. The bacterial genome is highly condensed into a nucleoid structure. Here we present global analyses of the genome spatial organization for two M-NM-3-proteobacteria: Escherichia coli and Pseudomonas aeruginosa by Genome Conformation Capture. Long distance interactions occurred within the E. coli and P. aeruginosa nucleoids with frequencies that were affected by growth condition and gene dosage. Spatial clustering of genes that are either up or down-regulated depended on the environmental signals, indicating a non-random functional organization of the nucleoid. The largest changes in gene expression upon amino acid starvation occurred in genes that participate in long-range interactions. These genes remained highly spatially clustered when transcript levels decreased. Environment specific interactions were related to DNA motifs but did not correlate with binding sites for nucleoid associated proteins. Overall we identify spatial organization as a significant factor in bacterial gene regulation and suggest that the prokaryotic operon is not simply a linear entity. 4 samples grown in LB; 4 samples treated with SHX. For Genome Conformation Capture (GCC) analyses, E. coli strains were recovered from -80M-BM-0C on LB agar (2%) plates for 24 hours. LB medium (3ml) starter cultures were inoculated and grown (37M-BM-0C, 220rpm, 16h). The Optical Density (OD600) of cultures was measured and used to inoculate LB test cultures to an OD600 of ~0.02. The test cultures were grown (37M-BM-0C, 220rpm) until the OD600 reached ~0.25 and then harvested. For the serine hydroximate (SHX) treated samples the cultures were treated with SHX (500 M-BM-5g/ml, 30 min) before harvesting. E. coli chromatin was prepared according to Rodley et al. (2009)with the following modifications. A total of 5*109 cells were cross-linked with formaldehyde (1% final v/v, 20min, RT) and then quenched with glycine (125mM final, 10min). Cells were collected by centrifugation (4000rpm, 15min, 4M-BM-0C), washed twice (1% PBS, 1% TritonX-100, 5ml/50ml culture) and pelleted (4000rpm, 15min, 4M-BM-0C). Cell pellets were suspended in 800M-NM-<l of B1 lysis buffer (10mM Tris pH 8.0, 50mM NaCl, 10mM EDTA, 20% (w/v) sucrose, 1mg/ml lysozyme) and incubated (37M-BM-0C, 30min). 800M-NM-<l of B2 lysis buffer (200mM Tris pH 8.0, 600mM NaCl, 4% TritonX-100, 1 protease inhibitor tablet (Roche) per 10ml of buffer added just before use) was gently added, mixed by inversion 3-4 times and incubated (37M-BM-0C, 10min). The cell lysate was centrifuged (21,500g, 20min, 4M-BM-0C) and the supernatant decanted. The chromatin was washed once with 1ml of chromatin digestion buffer (10mM Tris-HCl pH 8.0, 5mM MgCl2, 0.1% TritonX-100) by inverting the tube 3-4 times and centrifuged (21,500g, 20min, 4M-BM-0C). The supernatant was decanted and the chromatin pellet was suspended in 500M-NM-<l chromatin digestion buffer. Chromatin samples were aliquoted into 10 sets of 5*108 cells. Samples were digested with HhaI (100U, New England Biolabs). Following digestion a ligation control was added and the samples were ligated with T4 DNA ligase (20U, Invitrogen). Following ligation, cross-links were removed in the presence of proteinase K (0.45U, Fermentas). RNA was removed and pUC19 plasmid (27.4pg/2ml) was added as a sequencing control prior to three extractions with 1:1 Phenol:Chloroform. DNA was column purified (Zymo, DNA clean and concentratorTM-5 kit) according to the manufacturerM-bM-^@M-^Ys instructions and eluted in milliQ H2O before combining for sequencing. 3M-NM-<g of purified DNA was sent for pared-end sequencing at the ATC sequencing facility (Rockville, MD, USA) on an Illumina Hi-Seq. External ligation controls were produced by PCR amplification of short regions from the Lambda phage genome and the pRS426. Primers (Table S4 were designed to include an HhaI site at one end of the final product. PCR products were purified using a PCR purification kit (Qiagen), digested with HhaI (4U, 37M-BM-0C, 2h) and purified again. Purified, digested PCR products were introduced into the GCC samples at a 1:1 ratio with the number of genomes prior to the ligation step during GCC preparation. The pRS426 fragment was introduced into the exponential phase (LB grown) samples and resulted in 220 separate ligation events with HhaI restriction fragments on the genome. The Lambda phage fragment was introduced into the SHX treated samples and resulted in 2 ligation events with HhaI fragments on the genome. Network assembly was performed using the Topography suite v1.19. GCC networks were constructed from 100bp (E. coli) paired end Illumina Genome Analyser sequence reads. Topography uses the SOAP algorithm to position PE tags and single ends which contain a HhaI (E. coli) restriction site onto the E. coli (NC_000913) reference genomes, respectively. Each reference genome also contained the pUC19 (SYNPUC19CV) sequence. For the E. coli alignment the sequences of the pRS426 plasmid and Lambda phage ligation controls were also included in the file to which the sequences were aligned. No mismatches or unassigned bases (N) were allowed during positioning. Except where indicated, bioinformatics and statistical analyses were performed on interactions in which the sequence reads were able to be mapped uniquely onto the reference genome and were above the FDR cut-off value (5). All bioinformatics analysis was perfumed using in house Perl scripts.
2013-05-02 | E-GEOD-40603 | biostudies-arrayexpress