A Potential New Way to Treat Some of the Most Common Blinding Diseases (image)
Caption
Vascular endothelial growth factor (VEGF)/tumor necrosis factor (TNF) induces retinal permeability and tight junction loss at the cell border. Rats were injected intravitreally with vehicle [0.1 percent bovine serum albumin (BSA) in phosphate-buffered saline (PBS)] or VEGF/TNF (50/10 ng). A: Fluorescent angiography using Micron III was performed at 5 hours after intravitreal cytokine injection, followed by intravenous injection of fluorescein isothiocyanate--conjugated (FITC) BSA (100 mg/kg body weight) and 10-minute circulation. Generally increased retinal fluorescence, with clear changes at the optic nerve head, is readily observed (Micron III). After angiography, retinas were dissected and flat mounted for microscopic visualization, where vascular leak from capillaries was readily observed (microscope). B: Flat-mount retinas were analyzed for immunoreactivity of occludin (Occ) and zonula occludens protein 1 (ZO-1). Loss of occludin and ZO-1 immunostaining at endothelial cell borders was observed in the VEGF/TNF-treated retinas. C: aPKC ζ inhibitor (aPKC ζ-I) blocks VEGF/TNF-induced permeability. Rats were intravitreally injected with vehicle (0.1 percent BSA in PBS), VEGF/TNF (50/10 ng), or indicated dose of aPKC ζ-I in combination with VEGF/TNF. At 3 hours after intravitreal injection, animals received an i.v. injection of FITC-BSA (100 mg/kg body weight). Two hours later, animals were perfused with warm saline and retinas were removed for quantification of FITC-BSA accumulation. Results are expressed relative to vehicle control. Differences between groups were analyzed by analysis of variance with Tukey's post hoc test. Data are expressed as means ± SEM (C). *P < 0.05, ****P < 0.0001. Original magnification, x630 (A, right column, and B).
Credit
The American Journal of Pathology
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