title: Seahorse Xfe 24 Extracellular Flux Analyzer-Based Analysis of Cellular Respiration in Caenorhabditis elegans.
abstract: Mitochondria are critical for their role in ATP production as well as multiple nonenergetic functions, and mitochondrial dysfunction is causal in myriad human diseases. Less well appreciated is the fact that mitochondria integrate environmental and intercellular as well as intracellular signals to modulate function. Because mitochondria function in an organismal milieu, there is need for assays capable of rapidly assessing mitochondrial health in vivo. Here, using the Seahorse XF(e) 24 Extracellular Flux Analyzer and the pharmacological inhibitors dicyclohexylcarbodiimide (DCCD, ATP synthase inhibitor), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, mitochondrial uncoupler), and sodium azide (cytochrome c oxidase inhibitor), we describe how to obtain in vivo measurements of the fundamental parameters [basal oxygen consumption rate (OCR), ATP-linked respiration, maximal OCR, spare respiratory capacity, and proton leak] of the mitochondrial respiratory chain in the model organism Caenorhabditis elegans.
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, North Carolina.
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Luz
authors.affiliation: Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina.
authors.forename: Latasha L
authors.initials: LL
authors.lastname: Smith
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, North Carolina.
authors.forename: John P
authors.initials: JP
authors.lastname: Rooney
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; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D017173:Caenorhabditis elegans / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000031:analogs & derivatives / Q000494:pharmacology; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D025261:Mitochondrial Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D010101:Oxygen Consumption / Q000502:physiology*; D019810:Sodium Azide / Q000494:pharmacology
chemical_list: C108897:carbonylcyanide 4-trifluoromethoxyphenylhydrazone; D004024:Dicyclohexylcarbodiimide; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000255:Adenosine Triphosphate; D019810:Sodium Azide; D003576:Electron Transport Complex IV; D025261:Mitochondrial Proton-Translocating ATPases
keywords: Caenorhabditis elegans; Seahorse XFe24; mitochondrial respiration; mitochondrial toxicity