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Curr Protoc Toxicol. 2015 Nov 2;66:25.7.1-15. doi: 10.1002/0471140856.tx2507s66.

Seahorse Xfe 24 Extracellular Flux Analyzer-Based Analysis of Cellular Respiration in Caenorhabditis elegans.

Author information

1
Nicholas School of the Environment, Duke University, Durham, North Carolina.
2
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina.

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.

KEYWORDS:

Caenorhabditis elegans; Seahorse XFe24; mitochondrial respiration; mitochondrial toxicity

PMID:
26523474
PMCID:
PMC4632645
DOI:
10.1002/0471140856.tx2507s66
[Indexed for MEDLINE]
Free PMC Article

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