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A naphthalimide-based and Golgi-targetable fluorescence probe for quantifying hypochlorous acid.
Shu-Zhen Liu1, Jun-Hong Xu2, Qiu-Juan Ma1
1School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, PR China.
Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
|October 20, 2022
Summary
A new fluorescent probe detects hypochlorous acid (HOCl) in the Golgi apparatus. This probe offers a sensitive and selective method for imaging HOCl levels in living cells, aiding research into related diseases.
Area of Science:
- Biochemistry
- Cell Biology
- Chemical Biology
Background:
- The Golgi apparatus is crucial for cellular function, but stress can lead to excess reactive oxygen species (ROS).
- Hypochlorous acid (HOCl), a potent ROS, can cause Golgi dysfunction and is linked to neurodegenerative and autoimmune diseases.
- Current methods for detecting HOCl in the Golgi are limited, hindering disease research.
Purpose of the Study:
- To develop a novel fluorescent probe for specific detection and imaging of hypochlorous acid (HOCl) within the Golgi apparatus.
- To characterize the probe's sensitivity, selectivity, and performance in live-cell imaging.
- To establish a tool for investigating the role of Golgi-localized HOCl in cellular stress and disease.
Main Methods:
- Synthesis of a naphthalimide-based fluorescent probe incorporating phenylsulfonamide and dimethylthiocarbamate moieties.
- Investigation of the probe's fluorescence response to HOCl via intramolecular charge transfer (ICT) mechanisms.
- Evaluation of the probe's detection limit, selectivity, working pH range, and biocompatibility.
- Application of the probe for real-time imaging of Golgi HOCl in live Hela cells.
Main Results:
- The developed probe exhibits weak fluorescence in the absence of HOCl due to hindered ICT.
- Upon HOCl presence, ICT is initiated, resulting in strong green fluorescence.
- A linear correlation between fluorescence intensity and HOCl concentration (5.0 × 10⁻⁷ to 1.0 × 10⁻⁵ mol·L⁻¹) was observed.
- A low detection limit of 5.7 × 10⁻⁸ mol·L⁻¹ for HOCl was achieved.
- The probe demonstrated rapid response, high selectivity, broad pH tolerance, and good biocompatibility.
- Successful imaging of Golgi HOCl in live Hela cells was accomplished.
Conclusions:
- A novel, highly sensitive, and selective fluorescent probe for detecting hypochlorous acid in the Golgi apparatus has been successfully developed.
- The probe's ability to image intracellular HOCl in live cells opens avenues for studying Golgi-related diseases.
- This tool holds potential for investigating the role of endogenous and exogenous HOCl in cellular processes and disease pathogenesis.
