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Updated: Jan 17, 2026
Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
Published on: January 24, 2017
Rational Design of a Novel Dual-Functional Fluorescent Probe for Simultaneous Monitoring and Imaging of Mitochondrial
Xin Zhou1, Yuhan Luo1, Shixiong Jin1
1Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, International Innovation Center for Forest Chemicals and Materials, College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China.
Abstract:
Drug-induced liver injury (DILI) poses a terrible threat to human health. Carbon monoxide (CO) is documented to have hepatoprotective effects by relieving hepatic oxidative stress caused by DILI, and adenosine-5'-triphosphate (ATP) has been considered a vital biomarker for reflecting mitochondrial energy metabolism in DILI. However, the dynamic fluctuations and cooperative actions of CO and ATP in DILI remain unclear. Herein, a new fluorescent probe CCRD for the simultaneous detection of CO and ATP fluctuations in two well-separated emission channels without signal crosstalk was constructed. The nitro group on the coumarin unit of CCRD after reacting with CO, was selectively reduced to its amino form, affording a distinct blue emission at 448 nm. By comparison, the coordination of ATP with CCRD caused the ring opening of the rhodamine moiety, generating an intense red emission at 584 nm. The probe CCRD enabled dual-channel imaging of endogenous and exogenous CO and ATP within HepG2 cells. Furthermore, the synergistic dynamic changes of CO bursts and ATP consumption in live cells and mouse livers triggered by DILI were monitored for the first time by using CCRD. This work not only realizes the bidirectional channel imaging of CO and ATP in vivo and in vitro but also provides a powerful tool for the early diagnosis and therapy of DILI.
Insights
Researchers developed a novel fluorescent probe, CCRD, to simultaneously track carbon monoxide (CO) and adenosine-5'-triphosphate (ATP) in drug-induced liver injury (DILI). This tool offers new insights into DILI mechanisms and potential early diagnosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Medical Diagnostics
Background:
- Drug-induced liver injury (DILI) is a significant health concern.
- Carbon monoxide (CO) exhibits hepatoprotective effects against DILI by mitigating oxidative stress.
- Adenosine-5 -triphosphate (ATP) is a key biomarker for mitochondrial energy metabolism in DILI, but its dynamic changes with CO remain unclear.
Purpose of the Study:
- To develop a novel fluorescent probe for simultaneous detection of CO and ATP.
- To investigate the dynamic fluctuations and cooperative actions of CO and ATP in DILI.
- To provide a tool for early diagnosis and therapy of DILI.
Main Methods:
- Construction of a new fluorescent probe (CCRD) with dual-channel emission.
- Selective reaction of CCRD with CO (blue emission) and ATP (red emission) without crosstalk.
- Dual-channel imaging of endogenous and exogenous CO and ATP in HepG2 cells and mouse livers.
Main Results:
- CCRD successfully detected CO and ATP fluctuations in separate emission channels.
- Dual-channel imaging of CO and ATP was achieved in live cells and mouse livers.
- Synergistic dynamic changes of CO bursts and ATP consumption during DILI were monitored for the first time.
Conclusions:
- The CCRD probe enables bidirectional channel imaging of CO and ATP in vitro and in vivo.
- This study reveals the dynamic interplay between CO and ATP in DILI.
- CCRD serves as a powerful tool for the early diagnosis and potential therapy of DILI.
