Construction of a novel NIR-emissive rhodamine derivative for monitoring mitochondrial viscosity in ferroptosis

Abstract

Ferroptosis, an iron-dependent programmed cell death mechanism, is mediated by distinct molecular pathways of lipid peroxidation caused by intracellular iron supplementation and glutathione synthesis inhibition that cause oxidative damage to the cell membrane. Monitoring viscosity changes of mitochondria is essential for a deeper understanding of ferroptosis, as mitochondria will be shrunk with increased membrane density and leading to drastic mitochondrial viscous changes during ferroptosis process. Thus, it is essential to explore novel and efficient fluorescent probes for monitoring viscosity in organisms. In this work, we designed and synthesized a mitochondria-targeting probe TJ-FRP for cellular viscosity measurement via fluorescence imaging method. To obtain this probe, we firstly developed a novel modifiable fluorescent π-extended xanthene dye TJ-FR by replacing the benzoic acid group with a strong electron-withdrawing perfluorobenzoic acid group at the 9-position of xanthene framework. The dye not only presents emission wavelength at 758 nm and a large stokes shift of 142 nm in water, but also the dye is low biotoxic, membrane permeable. By reaction with 4-aminobutyltriphenylphosphonium bromide, TJ-FR was converted to the mitochondria-targeting probe TJ-FRP. TJ-FRP was successfully applied for the imaging of viscosity in living cells. Especially, the probe can be applied for visualizing mitochondrial viscosity changes during various inducers-stimulated ferroptosis process in model cells. These findings suggest that this novel NIR fluorescent probe can serve as a powerful tool to monitor the viscosity in biological samples and may provide new insights for various diseases during ferroptosis.