A Systematic Design and Synthesis of PET-based Fluorescent Probes for Monitoring pH During Mitophagy

Mitochondria are the powerhouse of the cell and function at pH similar to 8.0. Dysfunctions of mitochondria, includes mitochondrial damage, leading to pH alteration. Hence, researchers aim to develop efficient pH probes for tracking mitochondrial pH dynamics. Herein, we developed a PET-based fluorescent probe for pH monitoring during mitochondrial dysfunctions. Three derivatives were synthesized with a variable spacer's length in pentacyclic pyridinium fluorophores (PM-C2, PM-C3, and PM-C6). An efficient electron transfers from the receptor (tertiary amine) was observed in the case of PM-C2 compared to the other two derivatives. This PET process was inhibited when tertiary amine was protonated in acidic pH. However, PM-C3 showed minimal fluorescence intensity at similar conditions and almost negligible change in case of PM-C6, suggesting poor PET process for both the derivatives. Furthermore, DFT/TD-DFT quantum chemical calculation well supported this optical phenomena and PET process. Biocompatible, photostable, and mitochondria-specific PM-C2 could monitor pH dynamics during mitochondrial damage which were engulfed by lysosome, also known as mitophagy. This mitophagy process were induced by rapamycin and starvation, which can be monitored by turn-on fluorescence enhancement. This process was further validated by tracking Parkin-protein translocation from cytoplasm to damaged mitochondria using our developed probe.

Automated summary

A PET-based fluorescent probe, PM-C2, was developed for monitoring pH dynamics during mitochondrial dysfunctions. Compared to the other derivatives, PM-C2 showed higher fluorescence intensity and stronger pH sensitivity, making it suitable for tracking mitochondrial damage-induced pH alterations. The probe demonstrated biocompatibility, photostability, and mitochondria-specific targeting. The research was rated 8 out of 10.