In Situ Observation of Lysosomal Hypobromous Acid Fluctuations in the Brain of Mice with Depression Phenotypes by Two-Photon Fluorescence Imaging

Excessive oxidative stress is the main cause of neurotransmitter metabolism disorder in the brain with depression. Lysosomal hypobromic acid (HOBr) is an important reactive oxygen species produced in oxidative stress. Its abnormal content can lead to macromolecular damage and neurodegenerative diseases. However, due to the high reactivity and low concentration of HOBr and the lack of in situ imaging methods, the role of HOBr in depression is not clear. Herein, based on the HOBr-initiated aromatic substitution of a tertiary amine, we developed a novel two-photon (TP) fluorescence probe (NH-HOBr) for real-time visual monitoring of trace HOBr in living systems. NH-HOBr introduces N-(2-aminoethyl)-morpholine as a new recognition receptor for HOBr and a targeting group for lysosomes. It not only has excellent selectivity compared with other biomolecules (including hypochlorous acid), fast response (≤5 s) and high sensitivity (LOD = 15 nM) but also realizes sensitive detection of HOBr in cells, zebrafish, and mice tissues. It is worth noting that the in situ TP fluorescence imaging of mouse brain reveals the positive correlation between HOBr content and depression phenotype for the first time, providing strong direct evidence for the relationship between oxidative stress and depression. This work can provide reference to further study depression and the pathological mechanism of HOBr. In addition, HOBr-initiated aromatic substitution of a tertiary amine provides a new idea for the construction of specific and sensitive HOBr probes.