期刊
THERANOSTICS
卷 9, 期 19, 页码 5672-5680出版社
IVYSPRING INT PUBL
DOI: 10.7150/thno.33867
关键词
Fluorescent imaging; fluorescent probe; autophagy; endothelial injury
资金
- National Key Research and Development Program of China [2016YFE0125400]
- National Natural Science Foundations of China [81473202, 81573411, 81225022, 21642007, 21778048]
- Zhejiang Provincial Natural Science Foundation of China [Z16H310003, R18H300001]
- National Science & Technology Major Project Key New Drug Creation and Manufacturing Program [2018ZX09711002]
Autophagy is a dynamic and complicated catabolic process. Imaging autophagic flux can clearly advance knowledge of its pathophysiology significance. While the most common way autophagy is imaged relies on fluorescent protein-based probes, this method requires substantial genetic manipulation that severely restricts the application. Small fluorescent probes capable of tracking autophagic flux with good spatiotemporal resolution are highly demanable. Methods: In this study, we developed a small-molecule fluorogenic probe (AFG-1) that facilitates real-time imaging of autophagic flux in both intact cells and live mice. AFG-1 is inspired by the cascading nitrosative and acidic microenvironments evolving during autophagy. It operates over two sequential steps. In the first step, AFG-1 responds to the up-regulated peroxynitrite at the initiation of autophagy by its diphenylamino group being oxidatively dearylated to yield a daughter probe. In the second step, the daughter probe responds to the acidic autolysosomes at the late stage of autophagy by being protonated. Results: This pathway-dependent mechanism has been confirmed first by sequentially sensing ONOO- and acid in aqueous solution, and then by imaging autophagic flux in live cells. Furthermore, AFG-1 has been successfully applied to visualize autophagic flux in real-time in live mice following brain ischemic injury, justifying its robustness. Conclusion: Due to the specificity, easy operation, and the dynamic information yielded, AFG-1 should serve as a potential tool to explore the roles of autophagy under various pathological settings.
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