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dc.contributor.authorBuczynski, Matthew W.en
dc.contributor.authorStephens, Daren L.en
dc.contributor.authorBowers-Gentry, Rebecca C.en
dc.contributor.authorGrkovich, Andrejen
dc.contributor.authorDeems, Raymond A.en
dc.contributor.authorDennis, Edward A.en
dc.description.abstractArachidonic acid is released by phospholipaseA2 and converted into hundreds of distinct bioactive mediators by a variety of cyclooxygenases (COX), lipoxygenases (LO), and cytochrome P450s. Because of the size and diversity of the eicosanoid class of signaling molecules produced, a thorough and systematic investigation of these biological processes requires the simultaneous quantitation of a large number of eicosanoids in a single analysis. We have developed a robust liquid chromatography/tandem mass spectrometry method that can identify and quantitate over 60 different eicosanoids in a single analysis, and we applied it to agonist stimulated RAW264.7 murine macrophages. Fifteen different eicosanoids produced through COX and 5-LO were detected either intracellularly or in the media following stimulation with 16 different agonists of Toll-like receptors (TLR), G protein-coupled receptors, and purinergic receptors. No significant differences in the COX metabolite profiles were detected using the different agonists; however, we determined that only agonists creating a sustained Ca22+ influx were capable of activating the 5-LO pathway in these cells. Synergy between Ca22+ and TLR pathways was detected and discovered to be independent of NF-κB-induced protein synthesis. This demonstrates that TLR induction of protein synthesis and priming for enhanced phospholipase A2-mediated eicosanoid production work through two distinct pathways.en
dc.format.extent22834 - 22847 page(s)en
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.titleTLR-4 and Sustained Calcium Agonists Synergistically Produce Eicosanoids Independent of Protein Synthesis in RAW264.7 Cellsen
dc.typeArticle - Refereeden
dc.description.versionPublished (Publication status)en
dc.contributor.departmentSchool of Neuroscienceen
dc.title.serialJournal of Biological Chemistryen
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Scienceen
pubs.organisational-group/Virginia Tech/Science/COS T&R Facultyen

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Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
License: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International