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Updated: May 16, 2025
Author Spotlight: Detection of Mitophagy in Caenorhabditis elegans and Mammalian Cells Using Organelle-Specific Dyes
Published on: May 19, 2023
Detection of mitophagy in live cells with indole derived near-infrared fluorogenic probes
Xuanyuan Wang1, Wen Chen1, Shuangling Liu2
1College of Chemistry and Material Science, Hengyang Normal University, Hengyang 421001, PR China.
Abstract:
Mitophagy is an indispensable cellular process that plays a crucial role in regulating mitochondrial quality control and cellular metabolism. Therefore, monitoring the changes in the mitochondrial and lysosomal microenvironment during the mitophagy process is extremely important. However, existing mitophagy probes only target changes in a single indicator (viscosity, pH value, or polarity) within the microenvironment, which may reduce the selectivity and accuracy of assessing mitophagy in complex biological settings. To address this, we have developed a dual-channel detection near-infrared (NIR) fluorescent probe (ADMI). In vitro analysis experiments have shown that ADMI not only responds to pH and activates the NIR fluorescence channel but also that the green fluorescence channel exhibits high sensitivity to changes in polarity. This dual-response mechanism probe enables dual fluorescent detection of pH and polarity, providing a highly promising tool for monitoring the microenvironment of mitophagy in living cells. Ultimately, we applied ADMI to real-time monitoring of mitophagy induced by starvation or rapamycin, during which the decrease in pH and polarity resulted in a red shift in wavelength and increased fluorescence. Additionally, ADMI was able to observe changes in mitochondria during ferroptosis. This probe may serve as a useful tool for imaging mitophagy in living cells.
Insights
Researchers developed a novel dual-channel fluorescent probe (ADMI) to monitor mitophagy. This tool accurately tracks changes in cellular microenvironment during mitophagy and ferroptosis in living cells.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Imaging
Background:
- Mitophagy is essential for mitochondrial quality control and cellular metabolism.
- Monitoring the mitochondrial and lysosomal microenvironment during mitophagy is crucial.
- Existing probes lack selectivity and accuracy due to single-indicator detection.
Purpose of the Study:
- To develop a novel probe for simultaneous detection of multiple microenvironmental parameters during mitophagy.
- To enhance the accuracy and selectivity of mitophagy assessment in complex biological systems.
- To enable real-time monitoring of mitophagy and related cellular events in living cells.
Main Methods:
- Development of a dual-channel near-infrared (NIR) fluorescent probe (ADMI).
- In vitro analysis of ADMI's response to pH and polarity changes.
- Application of ADMI for real-time imaging of mitophagy induced by starvation and rapamycin.
- Observation of mitochondrial changes during ferroptosis using ADMI.
Main Results:
- ADMI exhibits dual-response detection, correlating pH with NIR fluorescence and polarity with green fluorescence.
- The probe accurately monitors mitophagy-induced changes in pH and polarity, indicated by wavelength shifts and fluorescence changes.
- ADMI successfully visualized mitochondrial alterations during ferroptosis.
Conclusions:
- The dual-response ADMI probe offers a highly promising tool for dual fluorescent detection of pH and polarity.
- ADMI enables precise, real-time monitoring of the mitophagy microenvironment in living cells.
- This probe facilitates advanced imaging of mitophagy and mitochondrial dynamics in various cellular contexts.
