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Visualizing Peroxynitrite in Microvessels of the Brain with Stroke Using an Engineered Highly Specific Fluorescent
Jianhua Xiong1, Weiwei Wang1, Caixia Wang1
1Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials & Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules, College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, China.
Researchers developed a new fluorescent probe, Rd-PN2, for highly specific detection of peroxynitrite (ONOO⁻). This probe enables real-time, in vivo visualization of ONOO⁻ in cerebral microvessels during stroke, aiding diagnosis and therapy.
Area of Science:
- Biomedical Engineering
- Chemical Biology
- Neuroscience
Background:
- Stroke is a major cause of death and disability worldwide.
- Malfunction of reactive oxygen and nitrogen species (ROS/RNS) in cerebral microvessels is linked to stroke pathology.
- Selective in vivo monitoring of species like peroxynitrite (ONOO⁻) is crucial for stroke diagnosis and treatment.
Purpose of the Study:
- To develop a novel fluorescence probe for highly specific detection of ONOO⁻.
- To enable real-time, in vivo visualization of ONOO⁻ in cerebral microvessels during stroke.
- To investigate the utility of the probe in tracking endogenous ONOO⁻ in biological systems.
Main Methods:
- Engineering of an indoline-2,3-dione moiety to create the fluorescence probe Rd-PN2.
- Testing probe specificity against various ROS/RNS, including in the presence of high concentrations of interfering species.
- Evaluating probe sensitivity, reaction speed, and performance in tracking endogenous ONOO⁻ in living cells, zebrafish, and mice with stroke models using two-photon microscopy.
Main Results:
- The developed probe Rd-PN2 demonstrated highly specific response to ONOO⁻, even with coexisting ROS/RNS.
- Rd-PN2 exhibited high sensitivity and rapid response to ONOO⁻.
- The probe successfully visualized endogenously generated ONOO⁻ in living cells and zebrafish, and in cerebral microvessels of mice with ischemic and hemorrhagic strokes.
Conclusions:
- Rd-PN2 is a precisely modulated fluorescence probe for real-time in vivo visualization of ONOO⁻ production in cerebral microvessels.
- This probe facilitates accurate monitoring of ONOO⁻ during stroke events.
- The findings contribute to a better understanding of ONOO⁻ functions in stroke and related neurological conditions.
