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Updated: Jun 5, 2025
A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation
Published on: July 13, 2018
A Novel Method for Mitochondrial Membrane Potential Detection in Heart Tissue Following Ischemia-reperfusion in Mice
Chao Yin1, Chen-Xing Huang1, Le Pan1
1Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China.
Objective:
Myocardial ischemia-reperfusion (I/R) injury is associated with a significant reduction in the mitochondrial membrane potential (MMP, ΔΨm). Fluorescence-based assays are effective for labelling active mitochondria in living cells; their application in heart tissue, however, represents a challenge because of a low yield of viable cardiomyocytes after cardiac perfusion. This study aimed to examine a novel method for detecting the changes in the MMP of mouse heart tissue following I/R injury.
Methods:
The I/R model was established, which was characterized by distinct ischemic area and apoptosis in heart tissue. The MMP was detected via a confocal microscope after the ascending aorta was clamped and the mitochondrial probe solution (containing Mito-Tracker Deep Red FM) was perfused from the apex via a peristaltic pump.
Results:
This method enabled the distribution of the probe solution throughout the cardiac tissue via the coronary circulation. Fluorescence detection revealed that the MMP was profoundly reduced in both ischemic area and border area following I/R when compared with that in the sham group. There was no obvious difference in the MMP of the remote area between the I/R group and the sham group.
Conclusion:
This study presents a novel method for detecting the MMP in heart tissue, and this method will facilitate the evaluation of changes in the MMP in different regions following I/R.
Insights
This study introduces a new method to measure mitochondrial membrane potential (MMP) in mouse heart tissue after ischemia-reperfusion injury. The novel technique successfully detected reduced MMP in injured areas, aiding I/R injury assessment.
Area of Science:
- Cardiovascular Biology
- Mitochondrial Physiology
- Cellular Injury Mechanisms
Background:
- Myocardial ischemia-reperfusion (I/R) injury significantly reduces mitochondrial membrane potential (MMP, ΔΨm).
- Assessing MMP in heart tissue is challenging due to low cardiomyocyte yield after perfusion.
- Current methods struggle with effective MMP detection in intact cardiac tissue.
Purpose of the Study:
- To investigate a novel method for detecting changes in MMP in mouse heart tissue following I/R injury.
- To evaluate the feasibility of using fluorescence-based assays for MMP assessment in cardiac tissue.
Main Methods:
- Established an I/R injury model in mouse hearts.
- Perfused a mitochondrial probe (Mito-Tracker Deep Red FM) via the apex to assess MMP using confocal microscopy.
- Utilized ascending aorta clamping and a peristaltic pump for controlled probe delivery.
Main Results:
- The novel perfusion method successfully distributed the probe throughout the cardiac tissue via coronary circulation.
- A significant reduction in MMP was observed in the ischemic and border zones of I/R injured hearts compared to sham controls.
- No significant difference in MMP was found in the remote area between I/R and sham groups.
Conclusions:
- A novel and effective method for detecting MMP in mouse heart tissue was developed.
- This technique facilitates regional evaluation of MMP changes in response to I/R injury.
- The method holds promise for improved understanding and assessment of cardiac injury.
