Division of Ocular Neurodegeneration, Institute for Ophthalmic Research, Centre for Ophthalmology, University of Tuebingen, Tuebingen, Germany.
Invest Ophthalmol Vis Sci. 2010 Jan;51(1):493-7. doi: 10.1167/iovs.09-4003. Epub 2009 Sep 8.
To identify individual cone photoreceptors in a transgenic mouse line in vivo based on selective expression of green fluorescent protein (GFP) using cSLO (confocal scanning laser ophthalmoscopy) and to use this approach to monitor cone cell fate in mouse models of retinal degeneration.
Transgenic mice expressing GFP under the control of a red-green opsin promoter (RG-GFP mice) were analyzed in vivo with respect to GFP expression in cone cells using cSLO and functional integrity using electroretinography (ERG). Histology was performed to correlate the pattern of GFP expression with light microscopic data. Longitudinal monitoring of cone survival was evaluated in crossbreds of RG-GFP mice with cpfl1 and Rpe65(-/-) mutant mice, respectively.
The authors found that RG-GFP transgenic mice had a stable GFP expression that did not interfere with retinal function up to at least 3 months of age. Thus, a longitudinal analysis of cone degeneration in individual RG cpfl1 and RG Rpe65(-/-) cross-bred mice in vivo was successfully performed and demonstrated distinct time frames of cone survival in the particular mouse model.
Monitoring GFP expression in cone photoreceptor cells, such as in the RG-GFP mouse, is a promising in vivo approach for the analysis of cone survival in mice.
利用共聚焦扫描激光检眼镜（cSLO），基于绿色荧光蛋白（GFP）的选择性表达，在转基因鼠系中鉴定活体中的单个视锥光感受器，并利用该方法监测视网膜变性小鼠模型中的视锥细胞命运。
用 cSLO 分析在红色-绿色视蛋白启动子（RG-GFP 小鼠）控制下表达 GFP 的转基因小鼠的视锥细胞中 GFP 表达情况，并用电视网膜图（ERG）检测其功能完整性。进行组织学研究以将 GFP 表达模式与光镜数据相关联。分别用 RG-GFP 小鼠与 cpfl1 和 Rpe65(-/-) 突变小鼠杂交，对视锥细胞存活进行纵向监测。
作者发现 RG-GFP 转基因小鼠的 GFP 表达稳定，至少在 3 个月大时不会干扰视网膜功能。因此，成功地对个别 RG cpfl1 和 RG Rpe65(-/-) 杂交小鼠体内的视锥细胞变性进行了纵向分析，并证明了特定小鼠模型中视锥细胞存活的不同时间框架。
监测如 RG-GFP 小鼠中视锥光感受器细胞中的 GFP 表达，是分析小鼠视锥细胞存活的一种很有前途的活体方法。
