VALIDATION OF LASER-INDUCED CHOROIDAL NEOVASCULARIZATION (CNV) MOUSE MODEL.

BARCELONA, PABLO F.; SUBIRADA, PAULA V.; SADDI, NABIL; MARQUEZ, MARILYNAN; VAGLIENTI, MARIA V.; RIDANO, MAGALI E.; MARQUEZ, GABRIEL; PAZ, MARÍA C.; LUNA, JOSÉ D.; SÁNCHEZ, MARÍA C.

Mar del Plata

Congreso; Reunión Conjunta SAIC SAI SAFIS 2018; 2018

SAIC SAI SAFIS

Age-related macular degeneration (AMD) in its neovascular form isthe leading cause of vision loss among adults above the age of 55.In the present study, we validate an established procedure for inductionof choroidal neovascularization (CNV) in mice. This procedureconsists in the perforation of Bruch?s membrane by laser-inducedphotocoagulation, mimicking neovascular form of AMD.Thus, this study was performed in this model, in order to characterizethe neovascular process and its impact on the retinal functionalityand the inflammatory profile. Due to, we previously demonstratedthat α2M, and its receptor (LRP1) participate during retinal NV wealso focalized in the participation of α2M/LRP1 system.C57BL/6 mice, 3?6 months of age, previously anesthetized weretreated with four spots of argon green laser photocoagulation pereye. After 7 days of laser burn, we analyzed the NV on choroid/RPE flatmounts by isolectin B4 staining using confocal microscopy.Using specific 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 levels of α2Mand LRP1 were analyzed by WB and the inflammatory profile, asIL1β, IL6, and TNFα using qPCR assay. The retinal functionality wasassessed by scotopic ERG.We had able to standardize the size of the lesion. An important numberof ECs and pericyte was observed around the lesion. Microgliawas localized in central and peripheral area of the lesion. While α2Mand LRP1 showed increase level of expression on cells close to thelesion. Accompanied with high pro-inflammatory and pro-angiogenicfactors. The scotopic ERG a- and b-wave were decreased (p<0.05)in the eyes of this animals.In conclusion, we were able to reproduce and validate the CNVmodel in our laboratory, which offer the opportunity to evaluate theparticipation of α2M/LRP1 system.