Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe.

摘要: 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 imaged relies on fluorescent protein-based probes, this method requires substantial genetic manipulation that severely restricts application. Small probes capable tracking with good spatiotemporal resolution are highly demanable. Methods: In study, we developed small-molecule fluorogenic probe (AFG-1) facilitates real-time imaging in both intact cells live mice. AFG-1 inspired by cascading nitrosative acidic microenvironments evolving during autophagy. It operates over two sequential steps. first step, responds to up-regulated peroxynitrite at initiation diphenylamino group being oxidatively dearylated yield daughter probe. second autolysosomes late stage protonated. Results: This pathway-dependent mechanism has been confirmed sequentially sensing ONOO- acid aqueous solution, then cells. Furthermore, successfully applied visualize mice following brain ischemic injury, justifying robustness. Conclusion: Due specificity, easy operation, information yielded, should serve as potential tool explore roles under various pathological settings.