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Masahiko Sugimoto, Bela Anand-Apte; Regulation of Retinal Pigment Epithelium Tight Junctions Following Laser Photocoagulation. Invest. Ophthalmol. Vis. Sci. 2011;52(14):888.
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To examine the effect of laser photocoagulation (PC) on cell-cell junctions of retinal pigment epithelium (RPE) cells.
Human RPE cells (ARPE-19) grown to confluence on glass coverslips were photocoagulated using a 532-nm diode laser (532nm, 0.1 seconds, 75µm, 150mW). All analyses were conducted at 0, 24, 48 and 72 hours following PC. Tight junction (TJ) integrity was evaluated by western blot analysis of cell lysates and immunofluorescence microscopy of intact cells for expression and localization of TJ proteins ZO-1 and occludin. The conditioned medium of cells was analyzed for secretion of various angiogenic factors following PC.
PC induced disruption of TJs occurred in a time-dependent fashion along with concomitant secretion of epidermal growth factor (EGF). PC-induced phosphorylation of EGFR was observed by western blot analysis of whole-cell lysates. EGF receptor specific inhibitor (AG1478) could prevent the TJ disruption following PC.
TJ disruption in RPE cells following PC is mediated by EGF.
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