Retinal Ganglion Cells in Transgenic Mice Expressing GFP as an Index of RGC Damage under Anesthetized Mice

Abstract

Abstract: : Purpose: We generated transgenic (Tg) mice in which green fluorescent protein (GFP) was specifically expressed retinal ganglion cells (RGC) in retina. The goal of this study is to determine whether assessment of fluorescence intensity in this Tg mice can be used as an index of RGC damage in vivo. Methods:We created a transgene containing the 5' regulatory sequence of the mice thy-1.2 gene fused to the GFP coding sequence. This transgene was used to generate Tg mice in which RGCs were labeled GFP specifically. In this Tg mice, the right retinas were subjected to transient periods of ischemia of 60 minutes by elevation of intraocular pressure. 4,7,14 days after re-perfusion, we observed retinas by Scanning Laser Ophthalmoscope (SLO) in the same mice and measured fluorescence intensity. Results: This transgene was used to generate seven lines of PCR-positive founders. Three of the lines had bright green fluorescent by fluorescence microscopy. The GFP fills the entire cell. In two lines, almost RGCs were GFP positive, whereas a few RGCs were labeled in other line. The fluorescence intensity was decreased to 50% at the right retinas in14 days after transient ischemia to the opposite retina having been no change. Conclusions: It is thought that this Tg mice may become the valuation model of RGC death because we can observe change of RGCs death in the same anesthetized mice.

Keywords: cell death/apoptosis • imaging methods (CT, FA, ICG, MRI, OCT, RTA, S • retina