Anthony L. Luz, Cristina Lagido, Matthew D. Hirschey, Joel N. Meyer
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引用次数: 27
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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.
用表达荧光素酶的秀丽隐杆线虫在体内测定线粒体功能:氧化磷酸化、糖酵解和脂肪酸氧化对毒物诱导功能障碍的贡献
线粒体是许多药物和环境毒物的靶标;然而,毒物诱导的线粒体功能障碍如何促进人类疾病的进展仍然知之甚少。为了解决这个问题,需要开发能够快速评估线粒体功能的体内检测方法。在这里,我们使用模式生物秀丽隐杆线虫,描述了如何使用表达荧光素酶的ATP报告菌株,快速评估有毒物质暴露后电子传递链、糖酵解或脂肪酸氧化在能量代谢中的体内作用。毒物暴露后线粒体功能的改变可以通过抑制线粒体电子传递链、糖酵解或脂肪酸氧化来消耗稳态ATP水平来检测。短期抑制剂暴露后ATP的差异变化表明毒性诱导的抑制部位的改变。因为微孔板读取器是唯一需要的主要设备,这是一种非常容易获得的方法来研究体内毒物诱导的线粒体功能障碍。©2016 by John Wiley &儿子,Inc。
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