Riccardo Natoli1,2, Matt Rutar1, Yen-Zhen Lu1, Joshua A Chu-Tan1, Yuwei Chen1, Kartik Saxena1, Michele Madigan3,4, Krisztina Valter1,2, Jan M Provis1,2. 1. a John Curtin School of Medical Research, Australian National University , Canberra , Australia. 2. b ANU Medical School, The Australian National University , Canberra , Australia. 3. c School of Optometry and Vision Sciences, University of New South Wales , Sydney , Australia. 4. d The Save Sight Institute, University of Sydney , Sydney , Australia.
Abstract
PURPOSE: Light is a requirement for the function of photoreceptors in visual processing. However, prolonged light exposure can be toxic to photoreceptors, leading to increased reactive oxygen species (ROS), lipid peroxidation, and photoreceptor cell death. We used the 661W mouse cone photoreceptor-like cell line to study the effects of pyruvate in protecting these cells from light-induced toxicity. METHODS: 661W cells were exposed to 15,000 lux continuous bright light for 5 hours and incubated in Dulbecco's modified eagle medium (DMEM) with various concentrations of pyruvate. Following light damage, cells were assessed for changes in morphology, cell toxicity, viability, and ROS production. Mitochondrial respiration and anaerobic glycolysis were also assessed using a Seahorse Xfe96 extracellular flux analyzer. RESULTS: We found that cell death caused by light damage in 661W cells was dramatically reduced in the presence of pyruvate. Cells with pyruvate-supplemented media also showed attenuation of oxidative stress and maintained normal levels of ATP. We also found that alterations in the concentrations of pyruvate had no effect on mitochondrial respiration or glycolysis in light-damaged cells. CONCLUSIONS: Taken together, the results show that pyruvate is protective against light damage but does not alter the metabolic output of the cells, indicating an alternative role for pyruvate in reducing oxidative stress. Thus, sodium pyruvate is a possible candidate for the treatment against the oxidative stress component of retinal degenerations.
PURPOSE: Light is a requirement for the function of photoreceptors in visual processing. However, prolonged light exposure can be toxic to photoreceptors, leading to increased reactive oxygen species (ROS), lipid peroxidation, and photoreceptor cell death. We used the 661W mouse cone photoreceptor-like cell line to study the effects of pyruvate in protecting these cells from light-induced toxicity. METHODS: 661W cells were exposed to 15,000 lux continuous bright light for 5 hours and incubated in Dulbecco's modified eagle medium (DMEM) with various concentrations of pyruvate. Following light damage, cells were assessed for changes in morphology, cell toxicity, viability, and ROS production. Mitochondrial respiration and anaerobic glycolysis were also assessed using a Seahorse Xfe96 extracellular flux analyzer. RESULTS: We found that cell death caused by light damage in 661W cells was dramatically reduced in the presence of pyruvate. Cells with pyruvate-supplemented media also showed attenuation of oxidative stress and maintained normal levels of ATP. We also found that alterations in the concentrations of pyruvate had no effect on mitochondrial respiration or glycolysis in light-damaged cells. CONCLUSIONS: Taken together, the results show that pyruvate is protective against light damage but does not alter the metabolic output of the cells, indicating an alternative role for pyruvate in reducing oxidative stress. Thus, sodium pyruvate is a possible candidate for the treatment against the oxidative stress component of retinal degenerations.
Entities:
Keywords:
661W cells; light-damage and oxidative stress; photoreceptors; retinal degeneration
Authors: Nilisha Fernando; Josephine H C Wong; Shannon Das; Catherine Dietrich; Riemke Aggio-Bruce; Adrian V Cioanca; Yvette Wooff; Joshua A Chu-Tan; Ulrike Schumann; Chinh Ngo; Rohan W Essex; Camilla Dorian; Sarah A Robertson; Si Ming Man; Jan Provis; Riccardo Natoli Journal: Front Cell Dev Biol Date: 2020-06-26
Authors: Virginia Puente-Muñoz; José M Paredes; Sandra Resa; José Damaso Vílchez; Michal Zitnan; Delia Miguel; María Dolores Girón; Juan M Cuerva; Rafael Salto; Luis Crovetto Journal: Sci Rep Date: 2019-02-07 Impact factor: 4.379