Generation of a Transgenic Rabbit Model of Retinal Degeneration
Generation of a Transgenic Rabbit Model of Retinal Degeneration
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DOI：
10.1167/iovs.08-2863
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发表时间：
2009-03-01
影响因子：
4.4
通讯作者：
Terasaki, Hiroko
中科院分区：
文献类型：
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作者：
Kondo, Mineo;Sakai, Takao;Terasaki, Hiroko
目的.目的建立转基因（Tg）兔视网膜变性模型，并通过组织学和电生理学研究其变性模式。通过BAC转基因产生视紫红质Pro347 Leu Tg兔。通过Southern印迹分析鉴定Tg兔，并通过定量RT-PCR测量表达水平。视网膜组织学检查采用光镜、电镜和免疫组化。通过全视野视网膜电图（ERG）评估视网膜功能。产生了六个品系的Tg兔，并且具有较高表达水平的两个品系显示出视杆细胞显性进行性视网膜变性。视网膜组织学检查表明，光感受器的损失有明显的区域变化，中央视网膜比周边视网膜受到更严重的影响。转基因兔视网膜电图的特征表明，48周时视网膜电图的视杆成分减少到仅5%，而野生型在同一时间点的视锥成分保持在35%。视网膜超微结构显示，光感受器的细胞外间隙内聚集了大量的小泡。据作者所知，这是第一个进行性视网膜变性的兔模型。由于兔子有大眼睛，易于操作和繁殖，它们将为研究视网膜变性的病理生理学和新的治疗方法提供有用的动物模型。（Invest Ophthalmol维斯科学。2009;50：1371-1377）DOI：10.1167/iovs.08-2863
PURPOSE. To generate a transgenic (Tg) rabbit model of retinal degeneration and to characterize the pattern of degeneration by using histology and electrophysiology.METHODS. Rhodopsin Pro347Leu Tg rabbits were generated by BAC transgenesis. Tg rabbits were identified by Southern blot analysis, and the expression levels were measured by quantitative RT-PCR. Retinal histology was examined by light and electron microscopy and immunohistochemistry. Retinal function was assessed by full-field electroretinograms (ERGs).RESULTS. Six lines of Tg rabbits were generated, and two lines with higher levels of expression showed rod-dominant progressive retinal degeneration. Retinal histology indicated a marked regional variation in the loss of photoreceptors with the central retina more severely affected than the peripheral retina. The characteristics of the ERGs of transgenic rabbits indicated that the rod components of the ERGs were reduced to only 5% by 48 weeks, whereas the cone components remained at 35% in the wild-type at the same time point. The retinal ultrastructure of Tg rabbits showed a large number of small vesicles that accumulated in the extracellular space of the photoreceptors.CONCLUSIONS. To the best of the authors' knowledge, this is the first rabbit model of progressive retinal degeneration. Because rabbits have large eyes and are easy to handle and breed, they will provide a useful animal model for the study of the pathophysiology of and new treatments for retinal degeneration. (Invest Ophthalmol Vis Sci. 2009;50:1371-1377) DOI: 10.1167/iovs.08-2863