Measurements of the mitochondrial respiration and glycolytic activity in Drosophila embryonic cells
Measurements of the mitochondrial respiration and glycolytic activity in Drosophila embryonic cells
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果蝇胚胎细胞线粒体呼吸和糖酵解活性的测量
DOI：
10.1038/protex.2017.069
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Imai Yuzuru
中科院分区：
文献类型：
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作者：
Meng Hongrui;Yamashita Chikara;Hattori Nobutaka;Imai Yuzuru
果蝇（Drosophila melanogaster）是一种用于了解人类疾病的强大动物模型，因为这种微小的动物具有与人类疾病相关的简单但保守的细胞信号通路。使用果蝇的研究有助于理解线粒体疾病的发病机制，包括神经退行性疾病和代谢性疾病。在这里，我们提供了耗氧率（OCR）和细胞外酸化率（ECAR）的测量方案，这些方案已使用Seahorse XF e 24细胞外通量分析仪针对来自早期发育阶段胚胎的果蝇细胞进行了优化。这些方法需要具有同步发育阶段的受精卵，所述受精卵已被机械均质化，随后铺在微孔板上用于测定。利用这些方案，可以在果蝇胚胎细胞中在各种线粒体应激期间真实的测量OCR和ECAR，并且可以估计氧化磷酸化（OXPHOS）期间的电子通量。
The fruit fly (Drosophila melanogaster) is a powerful animal model used to understand human diseases because this tiny animal has simple but conserved cell signaling pathways that are associated with human diseases. Studies using Drosophila have contributed to the understanding of the pathogenesis of mitochondrial diseases, including neurodegenerative diseases and metabolic diseases. Here, we provide measurement protocols for the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) that have been optimized for Drosophila cells from early developmental stage embryos using the Seahorse XF e 24 Extracellular Flux Analyzer. These methods require fertilized eggs with a synchronized developmental stage that have been mechanically homogenized and subsequently plated on a microplate for the assay. With these protocols, the OCR and the ECAR can be measured in Drosophila embryonic cells during a variety of mitochondrial stresses in real time and the electron flux can be estimated during oxidative phosphorylation (OXPHOS).