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P.S. Bora, J. Sohn, J.M. Cruz, S. Kang, H.J. Kaplan, N.S. Bora; Complement Activation is Required in the Murine Model of Laser-induced Choroidal Neovascularization . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3940.
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Purpose: To investigate the role of the complement system in the mouse model of laser-induced choroidal neovascularization (CNV). Methods: CNV was induced by laser photocoagulation in the C57BL/6 mouse with the krypton red laser (0.05 sec., 100 uM, 250 mW). Three laser spots were placed in each eye of four different groups of mice (n=5): C57BL/6, C57BL/6 treated with Cobra Venom Factor (CVF), C3 deficient and wild-type control for C3 deficient. Mice were perfused with 1ml of 3% dextran (molecular weight 2 million Daltons) prior to being sacrificed on day 7. Their eyes were enucleated, fixed (1h in 10% phosphate buffered formalin) and RPE-choroidal-scleral flat mounts stained with an anti-elastin (Cy-3 conjugated) antibody. The incidence and size of CNV was determined by confocal microscopy. Flat mounts were also stained for C3 and membrane attack complex (MAC). The purified IgG fraction of antiserum to mouse C3 was used to stain for C3, while a rabbit antibody to mouse C9 was used to localize MAC. Results: Triplicate experiments revealed the following incidence of CNV: C57BL/6 (control) = 98%; CVF treated C57BL/6 = 5%; C3 deficient wild type (control) = 98 %; C3 deficient = 10%. Additionally, the CNV complex in each of the experimental groups was markedly smaller than that in the controls. Laser spots stained very strongly for C3 and MAC in C57BL/6 mice and wild-type controls for C3-deficient animals. In contrast, no staining for C3 and MAC was observed in the laser spots in CVF treated as well as C3-deficient mice. Conclusions: These results suggest that the presence and activation of the complement system is essential for the development of laser-induced choroidal angiogenesis in mice.
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