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LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism
Journal article   Open access   Peer reviewed

LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism

Andrew J Worth, Dylan M Marchione, Robert C Parry, Qingqing Wang, Kevin P Gillespie, Noelle N Saillant, Carrie Sims, Clementina Mesaros, Nathaniel W Snyder and Ian A Blair
Journal of Visualized Experiments, v 2016(110), pp e53941-e53941
2016
PMID: 27077278
url
https://doi.org/10.3791/53941View
Published, Version of Record (VoR) Open

Abstract

Environmental Sciences
Perturbed mitochondrial metabolism has received renewed interest as playing a causative role in a range of diseases. Probing alterations to metabolic pathways requires a model in which external factors can be well controlled, allowing for reproducible and meaningful results. Many studies employ transformed cellular models for these purposes; however, metabolic reprogramming that occurs in many cancer cell lines may introduce confounding variables. For this reason primary cells are desirable, though attaining adequate biomass for metabolic studies can be challenging. Here we show that human platelets can be utilized as a platform to carry out metabolic studies in combination with liquid chromatography-tandem mass spectrometry analysis. This approach is amenable to relative quantification and isotopic labeling to probe the activity of specific metabolic pathways. Availability of platelets from individual donors or from blood banks makes this model system applicable to clinical studies and feasible to scale up. Here we utilize isolated platelets to confirm previously identified compensatory metabolic shifts in response to the complex I inhibitor rotenone. More specifically, a decrease in glycolysis is accompanied by an increase in fatty acid oxidation to maintain acetyl-CoA levels. Our results show that platelets can be used as an easily accessible and medically relevant model to probe the effects of xenobiotics on cellular metabolism.

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Collaboration types
Domestic collaboration
Web of Science research areas
Multidisciplinary Sciences
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