Leung Christopher K S, Lindsey James D, Crowston Jonathan G, Ju Won-Kyu, Liu Qwan, Bartsch Dirk-Uwe, Weinreb Robert N
Hamilton Glaucoma Center, Department of Ophthalmology, University of California, San Diego, La Jolla, CA, United States.
J Neurosci Methods. 2008 Mar 15;168(2):475-8. doi: 10.1016/j.jneumeth.2007.10.018. Epub 2007 Nov 7.
Current methods for in vivo retinal ganglion cells (RGCs) imaging involve either retrograde or intravitreal injection of chemical or biological tracers, which are invasive and may require repeated injection for serial long-term assessment. We have developed a confocal scanning laser ophthalmoscope technique (blue-light CSLO or bCSLO) to image retinal ganglion cells (RGCs) in mice expressing cyan fluorescent protein under the control of a Thy-1 promoter. Fluorescent spots corresponding to CFP-expressing retinal ganglion cells were discernable with the bCSLO. 96.1+/-2.6% of CFP expressing cells also were retrograde labeled with DiI indicating the bCSLO imaged fluorescent spots are RGCs. The imaging of Thy-1 promoter-driven CFP expression in these mice could serve as a sensitive indicator to reflect the integrity of RGCs, and provides a non-invasive method for longitudinal study of the mechanism of RGC degeneration and the effect of neuroprotective agents.
目前用于体内视网膜神经节细胞（RGCs）成像的方法包括逆行或玻璃体内注射化学或生物示踪剂，这些方法具有侵入性，并且可能需要重复注射以进行连续的长期评估。我们开发了一种共焦扫描激光检眼镜技术（蓝光CSLO或bCSLO），用于对在Thy-1启动子控制下表达青色荧光蛋白的小鼠视网膜神经节细胞（RGCs）进行成像。使用bCSLO可以辨别出与表达CFP的视网膜神经节细胞相对应的荧光斑点。96.1±2.6%表达CFP的细胞也被DiI逆行标记，表明bCSLO成像的荧光斑点是RGCs。在这些小鼠中对Thy-1启动子驱动的CFP表达进行成像，可以作为反映RGCs完整性的敏感指标，并为纵向研究RGCs变性机制和神经保护剂的作用提供一种非侵入性方法。
