PURPOSE: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). METHODS: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 3-10 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. RESULTS: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGF corneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8 mm(2) vs. 0.11 mm(2); p < 0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. CONCLUSIONS: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders.
PURPOSE: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). METHODS: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 3-10 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. RESULTS: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGFcorneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8 mm(2) vs. 0.11 mm(2); p < 0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. CONCLUSIONS: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders.
Authors: Damien Bates; G Ian Taylor; Joe Minichiello; Peter Farlie; Adam Cichowitz; Nadine Watson; Michael Klagsbrun; Roni Mamluk; Donald F Newgreen Journal: Dev Biol Date: 2003-03-01 Impact factor: 3.582
Authors: S Holmes; A M Downs; A Fosberry; P D Hayes; D Michalovich; P Murdoch; K Moores; J Fox; K Deen; G Pettman; T Wattam; C Lewis Journal: Scand J Immunol Date: 2002-09 Impact factor: 3.487