網膜神経節細胞蛍光発色トランスジェニックマウスを用いた非侵襲的網膜神経節細胞死の評価法の確立

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Purpose: We generated transgenic (Tg) mice in which green fluorescent protein (GFP) was specifically expressed retinal ganglion cell (RGC) in retina. The goal of this study is to determine whether assessment of fluorescent intensity in this Tg mouse can be used as an index of RGC damage in vivo. Method: We created a transgene containing the 5' promotor sequence of the mouse thy-1.2 gene and green fluorescent protein coding sequence. This transgene was injected to mouse fertilized eggs to generate transgenic mice in which RGCs specifically express GFP. In the Tg mouse line, the retina was subjected to transient ischemia (45 min.) by elevating of intraocular pressure. Four, seven and fourteen days after blood reperfusion, we monitored the retinal fluorescence of the mouse in vivo using Scanning Laser Ophthalmoscope (SLO) to assess neurodegeneration. Result: A total of seven transgenic founders were obtained. The eyes in three of these lines exhibited bright green fluorescence under fluorescence microscopy. In a mouse Tg-line, GFP filled the entire cell layers in the retina. In the other two lines, RGCs were specifically GFP-positive, or photoreceptor cells were labeled. Using SLO, we continuously observed the change in fluorescent intensity in non-invasive techinique. The fluorescent intensity of the retinal image with SLO decreased to 40% on 7 days after ischemia-reperfusion. Accordingly, there was a significant correlation between changes in fluorescent intensity of the Tg mouse retinas and the ganglion cell number. Conclusion: I obtained the mouse Tg line that elicits green fluorescence from retinal ganglion cells. The mouse line will be useful to monitor the ganglion cell degeneration non-invasively.
2004-06-10

網膜神経節細胞蛍光発色トランスジェニックマウスを用いた非侵襲的網膜神経節細胞死の評価法の確立

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