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P.S. Bora, S. Kaliappan, Y. Wang, S. Kumar, H.J. Kaplan, N.S. Bora; Expression of Complement Regulatory Proteins During Laser–Induced Choroidal Neovascularization . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3018.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: We have shown that the complement system plays a central role in laser induced choroidal neovascularization (CNV). This study was undertaken to analyze the expression (both mRNA and protein) of complement regulatory proteins – namely Crry and CD59 – in CNV lesions during laser–induced CNV in mice. Methods: CNV was induced by laser photocoagulation in the C57BL/6 mouse with the krypton red laser (0.05 sec., 100 uM, 250 mW). Animals were divided into two groups. In group 1, mice received three laser spots to induce CNV. Animals in group 2 received cobra venom factor (CVF), two days prior to laser photocoagulation and everyday after laser treatment. Animals from each group (n=6/group) were sacrificed on days 1, 3 and 7 post–laser. RPE–choroidal–scleral tissue was pooled separately from CVF treated and CVF untreated mice for semiquantative RT–PCR and Western blot analysis for Crry and CD59. Results: Our results of semi–quantative RT–PCR and Western blot analysis demonstrated that the levels of Crry and CD59 transcripts as well as protein were unaltered and were similar to the constitutive levels on day 1. However, on day 3 Crry and CD59 transcripts and protein were down–regulated. The levels of Crry and CD59 transcripts and protein returned to the constitutive levels on day 7 post–laser treatment. In contrast, in CVF treated animals the levels of Crry and CD59 in CNV lesions remained at the constitutive levels and did not change at different time points after laser treatment. Conclusions: Our results provide strong evidence that the expression of complement regulatory proteins during laser–induced CNV is altered to facilitate complement activation which is needed for the development of laser–induced CNV.
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