Vitreal macrophages express VEGF in oxygen-induced retinopathy
Purpose: The possibility of vitreal macrophages playing an angiogenic role in oxygen-induced retinopathy (OIR) was investigated. Oxygen-induced retinopathy was produced in newborn animals with the purpose of modeling the proliferative phase of human retinopathy of prematurity (ROP). Materials and Methods: To produce OIR in neonatal mice, litters at postnatal day 7 were placed in 80-90% oxygen for a period of 5 days and then returned to room air. Pups were killed on days 7, 12, 15, 17 and 20 over the postnatal period and were perfusion-fixed using a saline wash-out, followed by 4% paraformaldehyde and then India Ink. Eyes were enucleated and either whole-mounted, or snap-frozen and cryosectioned. Immunostaining procedures were used to visualize macrophages and vascular endothelial growth factor (VEGF) protein. The primary antibodies used were anti-F4/80 and antimouse VEGF, respectively. Vitreal macrophages closely associated with the vitreo-retinal interface (within 25 孠of the inner limiting membrane) were counted. In situ hybridization procedures were used to analyse for the presence of VEGF mRNA transcript in vitreal macrophages. Results: Macrophage numbers were found to significantly increase (P< 0.05) in eyes from oxygen-treated animals compared with those from age-matched controls. A close spatial relationship was observed between macrophages and vitreal neovascular sprouts. In addition, vitreal macrophages were also found to transcribe and express VEGF in the oxygen-treated animals during the vasoproliferative phase. Conclusions: Our results raise the possibility that vitreal macrophages play a role in the pathogenesis of OIR and by inference, ROP.
Clinical and Experimental Ophthalmology
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