Validation of laser-induced choroidal neovascularization (CNV) mouse model

BARCELONA, P; SUBIRADA, PV; SADDI, TN; MARQUEZ, MA; VAGLIENTI, MV; RIDANO, ME; MARQUEZ, GE; PAZ, MC; LUNA, JD; SANCHEZ, MC

Mar del Plata

Congreso; LXIII Reunión de la Sociedad Argentina de Investigación Clínica; 2018

Sociedad Argentina de Investigación Clínica (SAIC)

Age-related macular degeneration (AMD) in its neovascular form is the leading cause of visionloss among adults above the age of 55. In the present study, we validate an established procedurefor induction of choroidal neovascularization (CNV) in mice. This procedure consists in theperforation of Bruch?s membrane by laser-induced photocoagulation, mimicking neovascular formof AMD. Thus, this study was performed in this model, in order to characterize the neovascularprocess and its impact on the retinal functionality and the inflammatory profile. Due to, wepreviously demonstrated that α2M, and its receptor (LRP1) participate during retinal NV we alsofocalized in the participation of α2M/LRP1 system.C57BL/6 mice, 3?6 months of age, previously anesthetized were treated with four spots of argongreen laser photocoagulation per eye. After 7 days of laser burn, we analyzed the NV on choroid?RPE flatmounts by isolectin B4 staining using confocal microscopy. At the same time, usingspecific cell markers we characterize cells in the lesion area: CD105 (ECs), NG2 (pericytes),F4/80 (microglia) by immunostaining, as wells as LRP1 cell distribution. In addition, the proteinlevels of α2M and LRP1 were analyzed by WB and the inflammatory profile, as IL1β, IL6, andTNFα using qPCR assay. The retinal functionality was assessed by scotopic ERG. We had ableto standardize the size of the lesion. An important number of ECs and pericyte was observedaround the lesion. Microglia was localized in central and peripheral area of the lesion. While α2Mand LRP1 showed increase protein level and high expression pattern on cells close to the lesion.Accompanied with high pro-inflammatory and pro-angiogenic factors. The scotopic ERG a- andb-wave were decreased (p<0.05) in the eyes of this animals respect control. In conclusion, wewere able to reproduce and validate the CNV model in our laboratory, which offer the opportunityto evaluate the participation of α2M/LRP1 system.