Normal vascular development in mice deficient in endothelial NO synthase: possible role of neuronal NO synthase.

作者：Mohamed Al‐Shabrawey, Azza B. El‐Remessy, Xiaolin Gu, Steven S. Brooks, Mohamed S. Hamed, Paul L. Huang, Ruth B. Caldwell · 发表于：PubMed · 年份：2003 · 被引用次数：44 · 研究领域：Nitric Oxide and Endothelin Effects、Retinal Diseases and Treatments、Barrier Structure and Function Studies

PURPOSE: Nitric oxide formation by nitric oxide synthase (NOS) has been implicated in vascular injury and retinal neovascularization during oxygen-induced retinopathy. However, the role of NOS in normal retinal vascular development and growth has not been studied. The purpose of these experiments was to characterize the expression of NOS in relation to vascular development and to determine the effect of deleting endothelial NOS (eNOS) on this process. METHODS: Retinal vascular development was analyzed in 150 eNOS+/+ and eNOS-/- mice ranging from 1 day to 6 months old by using a combination of morphometric and biochemical approaches. The pattern of vascular development was analyzed in retinal tissue sections and whole-mount preparations labeled with fluorescein-conjugated Griffonia simplicifolia lectin. Analysis of vascular density and arterial diameter were performed with the lectin-labeled whole-mounts using computer-assisted morphometry. NO production was quantified by measuring retinal levels of nitrate/nitrite accumulation using the Greiss reaction. Western blotting techniques with isoform-specific NOS antibodies were used to evaluate differences in levels of NOS protein expression. Retinal distribution of nNOS was characterized using nNOS immunocytochemistry and NADPH diaphorase histochemistry. RESULTS: These analyses showed that the rate and pattern of retinal vascular development in eNOS-/- mice were comparable with those in wild-type control mice. Measurement of vascu...