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Purpose: To evaluate the utility of a novel profluorescent nitroxide (PFN) probe that selectively detects superoxide radicals in live cells, as a reporter of retinal oxidative status in models of retinal metabolic challenge.Method: (i) Cultured RGC-5 cells were transformed with trichostatin A to stimulate a “neuronal-like” morphology. Following treatment with 0.01 μM methylester rhodamine nitroxide probe, cells were exposed to the mitochondrial complex III inhibitor, antimycin (0.01−100 μM) to stimulate superoxide production. Fluorescence (556 nm/590 nm) was quantified via flow cytometry with statistical analysis based on 10,000 cells. (ii) Following intraocular injection of the PFN (2 μM), unilateral acute retinal ischaemia was induced in rats by elevation of intraocular pressure (IOP, 120 mmHg 60 min). After restoration of normal IOP, retinal fluorescence was assessed during reperfusion with a Micron III rodent fundus camera.Results: Antimycin caused a dose-dependent decrease in fluorescence in non-transformed and TSA-treated RGC-5 cells. The PFN probe detected a significant (p |
