PURPOSE: To determine the role of connexin43 (Cx43) and gap junctional intercellular communication (GJIC) in the response of the human retinal pigment epithelial cell line ARPE-19 to oxidative stress. METHODS: ARPE-19 cells were treated with the chemical oxidant tert-butyl hydroperoxide (t-BOOH), and cell viability was assessed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. GJIC was evaluated by scrape loading/dye transfer and microinjection assays, and Cx43 expression was detected by Western blot and immunofluorescent staining combined with confocal microscopy analysis. Retroviral infection of ARPE-19 cells with shRNA vectors targeting Cx43 or vectors encoding Cx43, Cx26, and a disease-linked dominant negative Cx43 mutant (G21R) were used, and the effect on cell viability was assessed. RESULTS: t-BOOH-induced ARPE-19 cell death was correlated with reductions in GJIC and in the total level of Cx43 protein expression. Overexpression of Cx26 and Cx43 increased the viability of oxidant-treated ARPE-19 cells. Conversely, shRNA knockdown of Cx43, expression of a disease-linked dominant negative Cx43 mutant, and blocking GJIC with 18beta-glycyrrhetinic acid and flufenamic acid all increased t-BOOH-induced ARPE-19 cell death. CONCLUSIONS: Cx43-mediated protection of ARPE-19 cells from oxidative stress-induced death is dependent on functional Cx43 channels.
PURPOSE: To determine the role of connexin43 (Cx43) and gap junctional intercellular communication (GJIC) in the response of the human retinal pigment epithelial cell line ARPE-19 to oxidative stress. METHODS: ARPE-19 cells were treated with the chemical oxidant tert-butyl hydroperoxide (t-BOOH), and cell viability was assessed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. GJIC was evaluated by scrape loading/dye transfer and microinjection assays, and Cx43 expression was detected by Western blot and immunofluorescent staining combined with confocal microscopy analysis. Retroviral infection of ARPE-19 cells with shRNA vectors targeting Cx43 or vectors encoding Cx43, Cx26, and a disease-linked dominant negative Cx43 mutant (G21R) were used, and the effect on cell viability was assessed. RESULTS:t-BOOH-induced ARPE-19 cell death was correlated with reductions in GJIC and in the total level of Cx43 protein expression. Overexpression of Cx26 and Cx43 increased the viability of oxidant-treated ARPE-19 cells. Conversely, shRNA knockdown of Cx43, expression of a disease-linked dominant negative Cx43 mutant, and blocking GJIC with 18beta-glycyrrhetinic acid and flufenamic acid all increased t-BOOH-induced ARPE-19 cell death. CONCLUSIONS:Cx43-mediated protection of ARPE-19 cells from oxidative stress-induced death is dependent on functional Cx43 channels.
Authors: Elisabeth Obert; Randy Strauss; Carlene Brandon; Christina Grek; Gautam Ghatnekar; Robert Gourdie; Bärbel Rohrer Journal: J Mol Med (Berl) Date: 2017-01-28 Impact factor: 4.599
Authors: Cady E Pocrnich; Qing Shao; Hong Liu; Mary M Feng; Sarah Harasym; Melissa Savage; Sarit Khimdas; Dale W Laird; Cindy M L Hutnik Journal: Graefes Arch Clin Exp Ophthalmol Date: 2011-12-03 Impact factor: 3.117
Authors: Yeri Kim; Jarred M Griffin; Mohd N Mat Nor; Jie Zhang; Peter S Freestone; Helen V Danesh-Meyer; Ilva D Rupenthal; Monica Acosta; Louise F B Nicholson; Simon J O'Carroll; Colin R Green Journal: Neurotherapeutics Date: 2017-10 Impact factor: 7.620
Authors: Abram Akopian; Tamas Atlasz; Feng Pan; Sze Wong; Yi Zhang; Béla Völgyi; David L Paul; Stewart A Bloomfield Journal: J Neurosci Date: 2014-08-06 Impact factor: 6.167
Authors: Hoa T Le; Wun Chey Sin; Shannon Lozinsky; John Bechberger; José Luis Vega; Xu Qiu Guo; Juan C Sáez; Christian C Naus Journal: J Biol Chem Date: 2013-12-03 Impact factor: 5.157