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TMT-based proteomics analysis of arachidonic acid (ARA) treated and control oocytes

ABSTRACT: Arachidonic acid (ARA) treated and control oocytes in 3 replicates were subjected to the 6-plex TMT labeling, HP-RP fractionation, and LC-MS/MS analysis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Oocyte

DISEASE(S): Disease Free

SUBMITTER: Xuejiang Guo

LAB HEAD: Xuejiang Guo

PROVIDER: PXD018753 | Pride | 2022-02-15

REPOSITORIES: Pride

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Publications

Characterization of Metabolic Patterns in Mouse Oocytes during Meiotic Maturation.

Li Ling L Zhu Shuai S Shu Wenjie W Guo Yueshuai Y Guan Yusheng Y Zeng Juan J Wang Haichao H Han Longsen L Zhang Jiaqi J Liu Xiaohui X Li Chunling C Hou Xiaojing X Gao Min M Ge Juan J Ren Chao C Zhang Hao H Schedl Tim T Guo Xuejiang X Chen Minjian M Wang Qiang Q

Molecular cell 20201016 3

Well-balanced and timed metabolism is essential for making a high-quality egg. However, the metabolic framework that supports oocyte development remains poorly understood. Here, we obtained the temporal metabolome profiles of mouse oocytes during in vivo maturation by isolating large number of cells at key stages. In parallel, quantitative proteomic analyses were conducted to bolster the metabolomic data, synergistically depicting the global metabolic patterns in oocytes. In particular, we disco ...[more]

PMID: 33068521

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TMT-based proteomics analysis of GV, GVBD and MII oocytes

Project description:Two replicates of GV, GVBD, and MII oocytes were subjected to the 6-plex TMT labeling, HP-RP fractionation, and LC-MS/MS analysis. Two labeling experiments were performed for the total four replicates of each stage of oocytes.

2022-02-15 | PXD018777 | Pride

Docosahexaenoic and Arachidonic Free Fatty Acids Exert Different Antitumor Activities on Colorectal Cancer Cells

Project description:Two PUFA, docosahexaenoic (DHA, 22: 6n3) and arachidonic acids (ARA, 20: 4n6), as well as their derivatives such as eicosanoids, regulate different activities, which include changes in receptor signaling, the composition of rafts, cell metabolism, and membrane structures. These also modify the function of transcription factors and their target genes, a key step essential for the function of FA-activated signaling. This work is focused to determine the antitumor actions of these compounds linked to the regulation of gene transcription on HT-29 colorectal cancer. For this, we performed antiproliferative antitumour assay, LDH breakage test, caspase-3 production, and proteome changes were assessed by SWATH-MS quantitative proteomics followed by pathway analysis in order to find out which molecular mechanisms were being affected. In all cases tested, DHA exercised antitumor actions to a higher extent than ARA, acting mainly by means of down-regulating most of the proteasome system particles, while ARA presented a heavy effect on all the six DNA replication helicase particles but did not affect proteasome. The results indicated that both DHA and ARA stopped colorectal cancer growth and proliferation, thus they represent the ideal candidates as chemopreventive agents.

2022-01-28 | PXD008653 | Pride

Gene Expression Analysis of Ara-C Resistance in AML

Project description:Using two independently derived murine BXH2 cell lines, Ara-C resistant derivatives were developed by exposure to increasing concentrations of Ara-C. Microarray analysis comparing the Ara-C resistant cells to their Ara-C sensitive parental cell lines identified potential genes involved in Ara-C resistance. Two highly Ara-C-resistant cell lines, B117H and B140H, were derived from Ara-C-sensitive parental cell lines, B117P and B140P. Variations in gene expression between these Ara-C-resistant and -sensitive sets were studied. Three replicates per cell line.

2010-03-07 | E-GEOD-18322 | biostudies-arrayexpress

Transcription profiling of two highly Ara-C resistant cell lines, B117H and B140H and two Ara-C sensitive parental cell lines, B117P and B140P to study altered gene expression in acute myeloid leukemia cell lines.

Project description:A microarray study of altered gene expression in acute myeloid leukemia cell lines Experiment Overall Design: Two highly Ara-C resistant cell lines, B117H and B140H were derived from Ara-C sensitive parental cell lines, B117P and B140P. Variations in gene expression between these resistant/sensitive sets were studied

2007-12-01 | E-GEOD-2788 | biostudies-arrayexpress

Transcriptome Sequencing (RNA-seq) of Ara-C Resistant Murine AML Cell Lines Identifies Mechanisms of Resistance

Project description:An RNA-seq study of altered gene expression and mutations in Ara-C resistant acute myeloid leukemia murine cell lines. The analysis of the RNA-seq data led to the identification of a large deletion within the Dck coding sequence of the B117H cell line, which produced an alternatively processed form of Dck mRNA. The RNA-seq analysis also identified the presence of an insertion mutation in Dck in the B140H cell line. The RNA-seq analysis also identified a number of significant expression changes which did not appear in a previous microarray analysis (GSE18322), as well as identified other mutations which may be contributing to Ara-C resistance. Two highly Ara-C resistant cell lines, B117H and B140H were derived from Ara-C sensitive parental cell lines, B117P and B140P. Variations in gene expression as well identification of acquired mutations between these Ara-C resistant/sensitive sets were studied using various RNA-seq analysis tools.

2014-08-13 | E-GEOD-47454 | biostudies-arrayexpress

Marker Frequency Analysis of Vibrio cholerae E7946 mutants (YAV20, AB14 and AB23)

Project description:YAV20 (E7946 loxP[dciA] SpR lacZ::cre ZeoR), AB14 (YAV20 inv[glmU-mioC]) and AB23 ( YAV20 ΔrecB::CmR) were created by natural transfromation using cognate plasmids. Cells were grown in the M( minimla media supplemented with fructose, with or without arabinose (ara). Genomic DNA was extracted with the Sigma GenElute® bacterial genomic DNA kit to generate a genomic library according to Illumina’s protocol. The libraries and the sequencing were performed by the High-throughput Sequencing facility of the I2BC (https://www.i2bc.paris-saclay.fr/sequencing/ng-sequencing/, CNRS, Gif-sur-Yvette, France). Genomic DNA libraries were made with the ‘Nextera DNA library preparation kit’ (Illumina) following the manufacturer’s recommendations.

2024-02-14 | E-MTAB-12729 | biostudies-arrayexpress

Gene expression profiling during Ara-C induced NB4 cell apoptosis

Project description:As the most important antimetabolite used to induce remission in children and adults with acute leukemia, cytosine arabinoside (Ara-C) is widely used in consolidation therapy of APL to reduce the relapse rate. In order to better understand the molecular mechanism underlying Ara-C induced cell early apoptosis, we examined the mRNA level changes of NB4 cells after Ara-C treatment over the 12 h time course using a human whole genome oligo-array. For each test and control sample at the 2, 4, 6, and 12 h time point, hybridization experiments were performed using a dye-swap strategy.

2011-10-14 | E-GEOD-32992 | biostudies-arrayexpress

Genes rapidly regulated by Ara-C and DNR treatment of HL-60 AML cells and role of NADPH-oxidase derived ROS in DNR-induced gene expression

Project description:Daunorubicin (DNR) and Cytarabin (Ara-C) are the main chemotherapeutic drugs used for Acute Myeloid Leukemias (AML) treatment. However, their mode of action is not well understood. To decipher if these drug would induce rapid transcriptional reprogramming, we treated HL-60 AML cell line with DNR or Ara-C for 3 hours and carried out a transcriptomic analyses. In addition, we had shown that Reactive Oxigen Species (ROS) produced by NADPH oxidases (NOX) are involved in DNR-induced gene expression. We therefore also performed a transcriptomic analyses in HL-60 cells treated with DNR and VAS2870, an NADPH oxidase inhibitor. HL-60 cells were treated or not for 3 hours with 2 ÂµM Ara-C, 1 ÂµM DNR or 1 ÂµM DNR + 20ÂµM VAS 2870. RNAs were purified from 3 independent experiments and used to probe Affymetrix Human Gene 2.0 ST Genechips

2017-12-31 | E-MTAB-4895 | biostudies-arrayexpress

Vitamin A and arachidonic acid altered the skeletal mineralization in Atlantic cod larvae (Gadus morhua) without any interactions on the transcriptional level

Project description:The main object of this study was to evaluate the impact of different levels of vitamin A (VA) and arachidonic acid (ARA) in relation to eicosapentaenoic acid (EPA) on mineralization and gene expression in Atlantic cod larvae (Gadus morhua). A two-way factorial design was used to identify possible interactions between the nutrients. First-feeding larvae were fed enriched rotifers from start-feeding until 29 days post hatch (dph). Larvae in four tanks were fed one of the following diets: control (EPA/ARA ratio: 15.8, 0.9 µg VA g-1 wet wt.), control+VA (EPA/ARA ratio: 15.8, 7.8 µg VA g-1 wet wt.), High ARA (EPA/ARA ratio: 0.9, 1.5 µg VA g-1 wet wt.) or High ARA +VA (EPA/ARA ratio: 0.9, 12.0 µg VA g-1 wet wt.). There was no difference in survival (33-40%) between the groups. Larvae fed High ARA+VA were shorter at 29 dph. The larvae fed High ARA+VA had significantly less mineralized bones, showing significant interaction effects between VA and ARA. Although transcriptomic analysis using a custom-made microarray did not reveal any interaction effects, it was found that high ARA diets altered the transcription of more genes than control +VA diets. Furthermore, blgap1 , blgap2 and col10a1 were all down-regulated in larvae fed High ARA-diets and to a lesser extent by Control +VA diet. Lower expression in the High ARA+VA diets compared to ARA and VA diets indicated an additive effect on mineralization. In conclusion, this study showed that the dietary increase in ARA and VA altered the skeletal metabolism during larval A. cod development, most likely through signaling pathways specific for each nutrient rather than an interaction. The present study also demonstrates that VA could affect the larval response to ARA, even within the accepted non-toxic/non-deficient range.

2015-03-17 | GSE66964 | GEO

Determination of allergen levels, isoforms and their hydroxyproline modifications among peanut genotypes by mass spectrometry

Project description:The recently published genome of peanut enables detailed molecular description of seed storage proteins, particularly allergens, which make up the peanut seed proteome. A detailed description of the allergenic proteome was produced using peanuts of different genetic backgrounds to increase the coverage of potentially expressed proteins and additionally allows for analysis of protein variability. Using relative quantification by mass spectrometry, minor variation of some allergenic proteins was observed but total levels of Ara h 1, 2, 3 and 6 were relatively consistent among genotypes. The isoform compositions of Ara h 1, 2 and 6 were similar. Three Ara h 3 isoforms were variable among genotypes and contributed to a large proportion of total Ara h 3 when present. The immunologically relevant amino acid modification hydroxyproline, previously thought to be present only on Ara h 2, was detected on Ara h 1 and 3 in consistent proportions given when the associated isoforms were present. The relevance of the data was evaluated in terms of food allergy regarding applications to reference materials, diagnostic and immunotherapeutic use, detection of peanuts in food, and risk assessment. The data can be used to design targeted mass spectrometry methods to permit analytical control of such materials.

2022-08-11 | PXD028086 | Pride

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