Study On Expression Phase And Localization Of NSA2in Choroidal Neovascularization Model In Mice Induced By Laser

Objective:(1) To establish choroidal neovascularization animal modelin mice.(2)Using RT-PCR,Western blotting and immunofluorescencestaining techniques to study the expression of Nop seven-associated2in eyetissues of choroidal neovascularization model in C57BL/6J mice. So as toprovide new exprimental and theoretical basis for exploration of thepathogenesis of wet AMD.Methods:(1) The532nm argon laser were used to induce CNV modelin C57BL/6J mice.(2) The CNV model eyes were taken at14days afterphotocoagulation to make choroidal spead, and CNV models were identifiedby the method of CD31antibody labeled with endothelial cells.(3) Theexpression pattern of NSA2mRNA and protein in CNV were examined byRT-PCR and Western blotting at1day,3days,5days,7days and14daysafter photocoagulation respectively.(4) The localization of NSA2protein inCNV models were detected by immunofluorescence staining at7days afterphotocoagulation.Results:(1) In this experiment, we use532nm argon laser induced a stable CNV aninal model in C57BL/6J mice successfully, can satisfy all theneeds of experiment in this project.(2) The PCR and Western blot resultsshow that NSA2mRNA and protein were weakly expressed in theretinal-choroid-sclera complex of normal adult C57BL/6J mice. After retinaphotocoagulation, the expression of NSA2mRNA and protein in CNVmodel eyes have significant time change rule. The expression intensity ofNSA2was gradully enhanced from1day through3days afterphotocoagulation, and reached the maximum at3days afterphotocoagulation, at14days it still slightly higher than normal levels，andthe difference had statistical significance (P<0.05).(3) Immunofluorescencestain results show that NSA2protein have strong expression in retinaplexiform layers in normal mouse.7days after laser photocoagulation,NSA2protein not only expressed in the retina plexiform, but also highexpressed in the CNV areas, the difference were statistically significant(P<0.05).Conclusion: Expression of NSA2were enhanced in the early period ofCNV formation, and it highly expressed in the CNV areas, so we infer thatNSA2may play an important role in the pathogenesis of CNV.