title: In Vivo Determination of Mitochondrial Function Using Luciferase-Expressing Caenorhabditis elegans: Contribution of Oxidative Phosphorylation, Glycolysis, and Fatty Acid Oxidation to Toxicant-Induced Dysfunction.
abstract: Mitochondria are a target of many drugs and environmental toxicants; however, how toxicant-induced mitochondrial dysfunction contributes to the progression of human disease remains poorly understood. To address this issue, in vivo assays capable of rapidly assessing mitochondrial function need to be developed. Here, using the model organism Caenorhabditis elegans, we describe how to rapidly assess the in vivo role of the electron transport chain, glycolysis, or fatty acid oxidation in energy metabolism following toxicant exposure, using a luciferase-expressing ATP reporter strain. Alterations in mitochondrial function subsequent to toxicant exposure are detected by depleting steady-state ATP levels with inhibitors of the mitochondrial electron transport chain, glycolysis, or fatty acid oxidation. Differential changes in ATP following short-term inhibitor exposure indicate toxicant-induced alterations at the site of inhibition. Because a microplate reader is the only major piece of equipment required, this is a highly accessible method for studying toxicant-induced mitochondrial dysfunction in vivo. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, North Carolina.
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Luz
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, Aberdeen, United Kingdom.
authors.forename: Cristina
authors.initials: C
authors.lastname: Lagido
authors.affiliation: Department of Medicine, Duke University, Durham, North Carolina.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Hirschey
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, North Carolina.
authors.forename: Joel N
authors.initials: JN
authors.lastname: Meyer
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects*; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D017930:Genes, Reporter / Q000187:drug effects; D006019:Glycolysis / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements; D008928:Mitochondria / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D023382:Toxicity Tests, Acute / Q000295:instrumentation / Q000379:methods*; D015262:Xenobiotics / Q000633:toxicity*
chemical_list: D005230:Fatty Acids, Nonesterified; D011993:Recombinant Fusion Proteins; D015262:Xenobiotics; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly
keywords: Caenorhabditis elegans; PE255; PE327; fatty acid oxidation; glycolysis; mitochondrial toxicity; oxidative phosphorylation