A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation

被引:23
|
作者
Chen, Guihao [1 ,2 ]
Yang, Yuejin [1 ,2 ]
Xu, Chuansheng [2 ,3 ]
Gao, Shuo [2 ,3 ]
机构
[1] Chinese Acad Med Sci, Natl Ctr Cardiovasc Dis, Fuwai Hosp, State Key Lab Cardiovasc Dis,Dept Cardiol, Beijing, Peoples R China
[2] Peking Union Med Coll, Beijing, Peoples R China
[3] Chinese Acad Med Sci, Natl Ctr Cardiovasc Dis, Fuwai Hosp, State Key Lab Cardiovasc Dis, Beijing, Peoples R China
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 137期
基金
中国国家自然科学基金;
关键词
Behavior; Issue; 137; Mitochondria! permeability transition pore; mitochondria! membrane potential; JC-1; assay; reperfusion injury; cardiac myocytes; flow cytometry; PERMEABILITY TRANSITION PORE; PROTECTS; CELLS; AUTOPHAGY; DEATH;
D O I
10.3791/57725
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Timely and efficient reperfusion of the occluded coronary artery is the best strategy for decreasing myocardial infarct size in patients with a ST-segment elevated myocardial infarction. However, reperfusion per se can result in further cardiomyocyte death, a phenomenon known as reperfusion injury. The opening of the mitochondria! permeability transition pore (mPTP), with the decrease of the mitochondria! membrane potential (MMP), or mitochondria! depolarization, is universally recognized as the final step of reperfusion injury and is responsible for mitochondria! and cardiomyocyte death. JC-1 is a lipophilic cationic dye that accumulates in mitochondria depending on the value of MMP. The higher the MMP is, the more JC-1 accumulates in the mitochondria. The increasing amounts of JC-1 in mitochondria can be reflected by a fluorescence emission shift from green (similar to 530 nm) to red (similar to 590 nm). Therefore, the reduction of the red/green fluorescence intensity ratio can indicate the depolarization of mitochondria. Here, we take advantage of JC-1 to measure MMP, or the opening of mPTP in human cardiac myocytes after hypoxia/reoxygenation, detected by flow cytometry.
引用
收藏
页数:7
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