Transcription profiling of Bos taurus mammary gland infected with E. coli reveals infection induces distinct local and systemic transcriptome responses in the mammary gland (Super Series)
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ABSTRACT: 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
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
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
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
Project description:Liver plays a profound role in the acute phase response (APR) observed in the early phase of acute bovine mastitis caused by Escherichia coli (E. coli). To gain an insight into the genes and pathways involved in hepatic APR of dairy cows we performed a global gene expression analysis of liver tissue sampled at different time points before and after intra-mammary (IM) exposure to E. coli lipopolysaccharide (LPS) treatment. Experiment Overall Design: Eight healthy, high yielding Holstein-Friesian dairy cows in their first lactation (9 to 12 weeks after calving) were chosen for this study. At time 0 the right front quarter was infused with 200 μg E. coli LPS dissolved in 10 ml 0.9% NaCl solution, the left front quarter serving as control was infused with 10 ml 0.9% NaCl solution. Liver biopsies were taken at â22, 3, 6, 9, 12 and 48 hours relative to LPS infusion in 4 cows, and also at â22, 9 and 48 hours in the remaining 4 cows. RNA from liver biopsies was isolated and biotin labeled cRNA was loaded onto the Affymetric GeneChip Bovine Genome Array. A control study using cows infused with 0.9% NaCl showed that there was no effect of taking the biopsy, neither in the clinical measurement nor in the expression of a selected subset of genes. Therefore, only samples taken from the LPS treated cows were measured for the gene expression using microarrays.
Project description:Previous studies have investigated the peptidomic changes occurring in cow milk during mastitis; however, these focused mainly on clinical mastitis, either spontaneous (Mansor et al., 2013) or induced by experimental infection (Thomas et al., 2016). Mansor and coworkers were the first to use mass spectrometry to demonstrate that several peptides found increased in milk from cows with clinical S. aureus or E. coli mastitis were mainly derived from aS1- and b-casein. In that study, 48 peptides were significantly different between the milks of healthy and mastitic cows. Non-mastitic samples were confirmed to be non-mastitic by having SCC <100,000 cells/mL (Mansor et al., 2013). Thomas and coworkers expanded the peptidomic repertoire in a study evaluating the kinetics of experimental S. uberis infection, and found signature peptides with potential as mastitis markers (Thomas et al., 2016). Only one study evaluated the milk peptidome in subclinical mastitis (Guerrero et al., 2015) demonstrating that even subclinical infections can cause significant increases in the total number of released peptides when compared to uninfected milk. However, neither the IMI agents nor the somatic cell counts were reported. With the aim of understanding high abundance protein and peptidomic changes due to subclinical CNS mastitis, to identify signature peptides with potential for subclinical mastitis detection, and to compare the proteomic and peptidomic findings with those reported in clinical mastitis, we investigated the influence of CNS IMI on high abundance milk proteins by SDS-PAGE and densitometric analysis, followed by a detailed characterization of the milk peptidome by means of high-performance liquid chromatography/tandem mass spectrometry and bioinformatic analysis.
Project description:This investigation reports a differential proteomic analysis of the secretome of six S. aureus strains belonging to two genotypes with opposite within-herd prevalence, GTB (high) and GTS (low), corresponding to sequence types (ST) 8 and 398, respectively. Total proteins released in the growth medium were subjected to high-resolution tandem mass spectrometry and differential analysis with Proteome Discoverer by label-free approach. Here, we reported both the characterization of secretome proteins and a panel of differential proteins specific of S. Aureus depending on high or low within-herd prevalence. GTB/ST8 (High) released more immunoglobulin-binding proteins, complement and antimicrobial peptide inhibitors, enterotoxins, and metabolic enzymes, while GTS/ST398 (Low) released more leukocidins, hemolysins, lipases, and peptidases. Furthermore, GTB/ST8 (High) released the von Willebrand factor protein, staphylokinase, and clumping factor B, while GTS/ST398 (Low) released the staphylococcal coagulase and clumping factor A. In conclusion, our results provide an in-depth characterization of secretome proteins of the three S. aureus GTB/ST8 and three GTS/ST398 strains with high and low within-herd prevalence. We describe the role of extracellular proteins in S. Aureus pathogenesis.
Project description:In recent years, due to the influence of climate change and rising sea temperature, the incidence of Vibrio alginolyticus infections is increasing, and becoming the second most common Vibrio species reported in human illness. Therefore, better understanding of the pathogenic mechanism of V. alginolyticus infection is urgently needed. Vvrr1 (Vibrio virulence regulatory RNA 1) is a new found ncRNA predicted to be closely related to the adhesion ability of V. alginolyticus through the previous RNA-seq. In this study, the target genes of Vvrr1 were fully screened and verified by constructing Vvrr1 over-expressed strains and proteome sequencing technology.
Project description:Corpus luteum (CL) is an ephemeral gland whose main function is to secrete progesterone required for the establishment and maintenance of pregnancy. It is very well established that development and maintenance of CL function in primates requires action of luteinizing hormone (LH) but the extent and mechanism by which LH contributes to the maintenance of CL function through out the luteal phase is not known. To study the nuclear actions mediated by LH, we evaluated global genomic changes in CL of monkeys treated with GnRH receptor antagonist to inhibit pituitary LH secretion. Affymetrix microarray analysis was performed on RNA samples from CL obtained from VEH or CET treated monkeys. Results demonstrate that LH regulates expression of a number of genes which might be important for maintenance of CL structure and function. Keywords: CL, LH, CET, gene expression A single s.c. injection of GnRH receptor antagonist, Cetrorelix (CET), administered at a dose of 150 µg/kg BW has been shown to induce luteolysis. For the purpose of microarray analysis, VEH (5.25% glucose) or CET (treatment) was injected to female monkeys on day 7 of luteal phase and CL collected at 24 h post treatment (n=3) was cut into small quarters and snap frozen in liquid nitrogen. To minimize between animal variations, CL for both VEH and CET treatments were collected from the same monkeys.
Project description:The benefit of treatment in mild to moderate cases of E. coli mastitis in dairy cows remains a topic of discussion. We investigated the effect of intramammary treatment with Cefapirin + Prednisolone compared to Cefapirin only on gene expression profiles in experimentally induced E. coli mastitis. Five midlactating Holstein Friesian cows were challenged with 100 CFU in 3 quarters and treated with Cefapirin only in one quarter and the combination of Cefaprirn and Prednisolne in another quarter at 4, 12, 24 and 36h post challenge. After 24h (n=2) or 48h (n=3) post challenge cows were sacrificed and mammary gland tissue was collected from each quarter. From each cow total RNA of one non challenged quarter, one challenged not treated quarter, on Cefapirin treated and one Cefapirin + Prednisolone treated quarter was subjected to microarray analysis. Data were analyzed separately for the two sample time points
Project description:Mastitis in dairy cows is one of the most costly and prevalent diseases affecting dairy cows world wide. Insight in the molecular regulation of the host immune response to an E. coli infection, could help to develop new strategies to prevent cattle from E. coli infection. Here we performed a gene-expression analysis from udder tissue exposed to a controlled E. coli infection at T=24h post infection (p.i.) representing the acute phase response and at T=192h p.i. representing a chronic stage. 49 Samples