Anal. Chem. 2021, 93, 2490−2499 Full text

Xianzhu Luo, Ziyi Cheng, Rui Wang,* and Fabiao Yu*



Epilepsy is a chronic neurodegenerative disease that has seriously threatened human health. Accumulating evidence

reveals that the pathological progression of epilepsy is closely related to peroxynitrite (ONOO−). Unfortunately, understanding the

physiological roles of ONOO− in epilepsy is still challenging due to the lack of powerful imaging probes for the determination of the

level of flfluctuations of ONOO−

in the epileptic brain. Herein, a near-infrared (NIR) two-photon (TP) flfluorescent probe

[dicyanomethylene-4H-pyran (DCM)−ONOO] is presented to trace ONOO− in living cells and in kainate (KA)-induced rat

epilepsy models with satisfactory sensitivity and selectivity. The probe is composed of a NIR TP DCM flfluorophore and a

recognition moiety diphenylphosphinamide. The phosphoramide bond of the probe is interrupted after reacting with ONOO− for

10 min, and then, the released amino groups emit strong flfluorescence due to the restoration of the intramolecular charge transfer

process. The probe can effffectively detect the changes of endogenous ONOO− with excellent temporal and spatial resolution in living

cells and in rat epileptic brain. The imaging results demonstrate that the increasing level of ONOO− is closely associated with

epilepsy and severe neuronal damage in the brain under KA stimulation. In addition, the low-dose resveratrol can effffectively inhibit

ONOO− overexpression and further relieve neuronal damage. With the assistance of TP flfluorescence imaging in the epileptic brain

tissue, we hypothesize that the abnormal levels of ONOO− may serve as a potential indicator for the diagnosis of epilepsy. The TP

flfluorescence imaging based on DCM−ONOO provides a great potential approach for understanding the epilepsy pathology and