Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes
Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes
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DOI：
10.1016/s0008-6363(00)00002-x
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发表时间：
2000-04-01
影响因子：
10.8
通讯作者：
Erusalimsky, JD
中科院分区：
文献类型：
--
作者：
Mathur, A;Hong, Y;Erusalimsky, JD
目标：线粒体膜电位（Δ psi m）的维持对于具有高能量需求的细胞如心肌细胞的正常性能和存活是基本的。因此，为了了解控制心肌细胞功能的分子机制，测量Δ psi m是必不可少的。在这里，我们评估了各种电位染料检测Δ psi m变化的能力。使用流式细胞术和共聚焦显微镜。方法：原代培养的乳鼠心肌细胞在Rho 123负载前后用线粒体解偶联剂处理。DiOC（6）（3）、CMXRos或JC-1，然后通过流式细胞术进行分析。光散射和Annexin V染色鉴定凋亡细胞。结果：所测试的四种电位染料能够区分活细胞和凋亡细胞。然而，只有JC-1能够检测到线粒体呼吸解偶联剂诱导的Delta psi m的崩溃。共聚焦显微镜分析证实，JC-1染色线粒体的电位依赖性的方式。相反。CMXRos染色心肌细胞，与Δ psi m的改变无关。结论：我们得出的结论是，JC-1是测量心肌细胞Delta psi m时使用的最佳染料。(C)2000由Elsevier Science B. V.出版，保留所有权利。
Objective: Maintenance of the mitochondrial membrane potential (Delta psi m) is fundamental for the normal performance and survival of cells such as cardiomyocytes, that have a high energy requirement. Measurement of Delta psi m is therefore essential in order to develop an understanding of the molecular mechanisms controlling cardiomyocyte function. Here We have evaluated various potentiometric dyes for their ability to detect alterations of Delta psi m. using flow cytometry and confocal microscopy. Methods: primary cultures of cardiomyocytes from neonate rats were treated with mitochondrial uncouplers before or after loading with Rho123. DiOC(6)(3), CMXRos or JC-1, and then analysed by flow cytometry. Apoptotic cells were identified by light scatter and Annexin V staining. Results: The four potentiometric dyes tested were able to discriminate between viable and apoptotic cells. However, only JC-1 was able to detect the collapse of Delta psi m induced by uncouplers of mitochondrial respiration. Confocal microscopic analysis confirmed that JC-1 stained mitochondria in a potential-dependent manner. In contrast. CMXRos stained cardiomyocytes irrespective of alterations in Delta psi m. Conclusions: We conclude that JC-1 is the optimal dye to use when measuring Delta psi m in cardiomyocytes. (C) 2000 Published by Elsevier Science B.V. All rights reserved.