Department of Applied Visual Science, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Analyst. 2019 Apr 8;144(8):2531-2540. doi: 10.1039/c8an02213e.
Mitochondrial activity is a widely used criterion to judge the metabolic condition of a living specimen. Numerous methods have been developed for related analyses, including the detection of O2 consumption, trans-membrane potential, and ATP production. In this study, we demonstrate that the redox state of cytochromes can serve as a sensitive mitochondrial activity indicator in glutamate-stressed neuronal cells. Mitochondrial dysfunction was detected by Raman imaging as early as 30 min after glutamate-stress induction. By comparing this result with other commonly used mitochondrial function assays, we found Raman imaging has a similar sensitivity to ATP production and trans-membrane potential assays. Other viability tests, such as MTT assay and ROS production tests, showed a slower response than our method. A thorough understanding of cytochrome dynamics with our new method will help establish Raman spectroscopy as a competitive clinical diagnosis tool for neurodegenerative diseases involving mitochondrial dysfunction.
线粒体活性是判断生物样本代谢状态的常用标准。已经开发出许多相关分析方法，包括检测 O2 消耗、跨膜电位和 ATP 生成。在这项研究中，我们证明细胞色素的氧化还原状态可以作为谷氨酸应激神经元细胞中线粒体活性的敏感指标。谷氨酸应激诱导后 30 分钟，通过拉曼成像即可检测到线粒体功能障碍。通过将这一结果与其他常用的线粒体功能测定方法进行比较，我们发现拉曼成像与 ATP 生成和跨膜电位测定具有相似的灵敏度。其他存活测试，如 MTT 测定和 ROS 生成测定，其反应速度比我们的方法慢。通过我们的新方法对细胞色素动力学进行深入了解，将有助于确立拉曼光谱作为涉及线粒体功能障碍的神经退行性疾病的一种有竞争力的临床诊断工具。
