Abstract
Purpose: :
To characterize the involvement of Semaphorin 7A in macrophage recruitment and corneal neovascularization
Methods: :
We generated anti-semaphorin 7A antibodies to detect the protein expression. Corneal fibroblast cells were cultured, stimulated with bFGF, immunostained with anti-Semaphorin 7A antibodies and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for days 3-10, corneal sections were imunostained with anti-semaphorin 7A antibodies. Mouse corneas were injected with naked semaphorin 7A DNA and vector control for 3, 7 and 10 days. Mouse corneas were photographed by slit lamp microscopy and areas of corneal neovascularization were calculated using ImageJ program. Mouse corneal sections also imunostained with anti-marcophage marker (F4/80) and VEGF-A.
Results: :
Our data showed an enhanced semaphoring 7A expression in bFGF stimulated cultured corneal fibroblast cells. bFGF corneal implantation also demonstrated an enhanced semaphorin 7A expression. Corneas injected with a Sema7A expression vector showed evidence of Sema7A protein expression and exhibited significant corneal NV compared to controls on day 10 (1.8 mm2 vs. 0.11 mm2; p<0.02). Immunolocalization of Sema7A DNA-injected corneas revealed macrophage recruitment and enhanced vascular endothelial growth factor-A expression
Conclusions: :
We demonstrated that Sema7A was expressed in vascularized corneas and showed proangiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal neovascularization and other angiogenesis-related disorders such as diabetic retinopathy, macular degeneration, and cancer.
Keywords: neovascularization • cornea: stroma and keratocytes • cornea: basic science