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K Fujisawa, C Spee, DR Hinton, SJ Ryan; Visualization of Vascular Endothelial Growth Factor Promoter Activity in Organ Cultured Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2758.
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Purpose: The aim of this study is to develop an in vivo detection system for vascular endothelial growth factor (VEGF) promoter activity in retina and observe the effects of various forms of stimulation. Method: The enucleated eyes of VEGF-green fluorescent protein (GFP) transgenic mice were used for this study. Each eye was cut at the equator. The posterior pole eyecups were placed in a 24-well culture dish and medium was added (neurobasal medium, B-27, L-glutamine, penicillin, streptomycin). Stimulation (hypoxia, VEGF, angiopoietin1, angiopoietin2, EphB4, ephrinB2 or nitric oxide) was applied on the second culture-day. Retinal GFP fluorescence was observed by confocal laser microscopy 1, 3, 6, 12 and 24 hours after stimulation. Results: VEGF promoter activity in retina was higher along veins than along arteries. Hypoxia and VEGF increased retinal VEGF promoter activity 12 hours after stimulation Conclusion: The regulation of VEGF promoter activity can be easily monitored using in vitro cultured VEGF-GFP retinas. This system is useful to observe transcription factor activity in vivo. Support:NIH grants EY03040 and EY01545 and grants from the Arnold and Mabel Backman Foundation and Research to Prevent Blindness. CR:N
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