Luz Anthony L, Smith Latasha L, Rooney John P, Meyer Joel N
Nicholas School of the Environment, Duke University, Durham, North Carolina.
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina.
Curr Protoc Toxicol. 2015 Nov 2;66:25.7.1-25.7.15. doi: 10.1002/0471140856.tx2507s66.
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.
线粒体对于其在ATP生成以及多种非能量功能方面的作用至关重要，线粒体功能障碍是众多人类疾病的病因。但人们较少认识到的一个事实是，线粒体整合环境信号、细胞间信号以及细胞内信号来调节功能。由于线粒体在生物体环境中发挥作用，因此需要能够在体内快速评估线粒体健康状况的检测方法。在此，我们使用海马XF（e）24细胞外通量分析仪以及药理学抑制剂二环己基碳二亚胺（DCCD，ATP合酶抑制剂）、羰基氰-对-三氟甲氧基苯腙（FCCP，线粒体解偶联剂）和叠氮化钠（细胞色素c氧化酶抑制剂），描述了如何在模式生物秀丽隐杆线虫中获得线粒体呼吸链基本参数[基础氧消耗率（OCR）、ATP关联呼吸、最大OCR、备用呼吸能力和质子泄漏]的体内测量值。
