Phenotypic characterization of P23H and S334ter rhodopsin transgenic rat models of inherited retinal degeneration.
Phenotypic characterization of P23H and S334ter rhodopsin transgenic rat models of inherited retinal degeneration.
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DOI：
10.1016/j.exer.2017.10.023
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发表时间：
2018-03
影响因子：
3.4
通讯作者：
Flannery JG
中科院分区：
文献类型：
--
作者：
LaVail MM;Nishikawa S;Steinberg RH;Naash MI;Duncan JL;Trautmann N;Matthes MT;Yasumura D;Lau-Villacorta C;Chen J;Peterson WM;Yang H;Flannery JG
我们在白化 Sprague-Dawley (SD) 大鼠中培育了 8 个表达两种不同视紫红质突变之一的转基因 (Tg) 大鼠系。在密码子 23 (P23H) 处用脯氨酸取代组氨酸产生了三个品系，这是美国最常见的常染色体显性视网膜色素变性形式。生成了 5 个品系，其终止密码子位于 334 位（S334ter），导致 C 端截短的视蛋白蛋白缺少最后 15 个氨基酸残基，并包含参与视紫红质失活的所有磷酸化位点，以及对视紫红质失活和运输很重要的末端 QVAPA 残基。这些模型中光感受器（PR）变性的速度与突变型与野生型视紫红质的比例成正比。这些模型已被广泛研究，但其表型的许多方面尚未得到描述。在这里，我们对 8 个 Tg 系进行了全面的研究，包括 PR 变性从发病到一岁的时间过程、光学和电子显微镜 (EM) 的视网膜结构、半球不对称性和视杆细胞和视锥细胞变性的梯度、视紫红质含量、基因剂量效应、假定小胶质细胞对外视网膜的快速激活和侵袭、视杆细胞外节盘脱落和视网膜色素上皮的吞噬作用(RPE) 和视网膜电图 (ERG) 的视网膜功能。注意到 PR 细胞死亡的双相性质，以及大鼠模型中缺乏损伤诱导的保护反应。电镜分析显示，在 S334ter 系 PR 外节变性的高峰期，亚微米囊泡结构在光感受器间空间中积累。这可能是由于消除了之前所见的运输共有结构域，就像其他缺乏 C 端 QVAPA 的视紫红质突变体一样。 8 个视紫红质 Tg 系已经并将继续成为遗传性视网膜变性实验研究中极其有用的模型。
We produced 8 lines of transgenic (Tg) rats expressing one of two different rhodopsin mutations in albino Sprague-Dawley (SD) rats. Three lines were generated with a proline to histidine substitution at codon 23 (P23H), the most common autosomal dominant form of retinitis pigmentosa in the United States. Five lines were generated with a termination codon at position 334 (S334ter), resulting in a C-terminal truncated opsin protein lacking the last 15 amino acid residues and containing all of the phosphorylation sites involved in rhodopsin deactivation, as well as the terminal QVAPA residues important for rhodopsin deactivation and trafficking. The rates of photoreceptor (PR) degeneration in these models vary in proportion to the ratio of mutant to wild-type rhodopsin. The models have been widely studied, but many aspects of their phenotypes have not been described. Here we present a comprehensive study of the 8 Tg lines, including the time course of PR degeneration from the onset to one year of age, retinal structure by light and electron microscopy (EM), hemispheric asymmetry and gradients of rod and cone degeneration, rhodopsin content, gene dosage effect, rapid activation and invasion of the outer retina by presumptive microglia, rod outer segment disc shedding and phagocytosis by the retinal pigmented epithelium (RPE), and retinal function by the electroretinogram (ERG). The biphasic nature of PR cell death was noted, as was the lack of an injury-induced protective response in the rat models. EM analysis revealed the accumulation of submicron vesicular structures in the interphotoreceptor space during the peak period of PR outer segment degeneration in the S334ter lines. This is likely due to the elimination of the trafficking consensus domain as seen before as with other rhodopsin mutants lacking the C-terminal QVAPA. The 8 rhodopsin Tg lines have been, and will continue to be, extremely useful models for the experimental study of inherited retinal degenerations.
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DOI：
10.1038/mt.2015.220
发表时间：
2016-03
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
Bakondi B;Lv W;Lu B;Jones MK;Tsai Y;Kim KJ;Levy R;Akhtar AA;Breunig JJ;Svendsen CN;Wang S
通讯作者：
Wang S
影响因子：
1.8
作者：
Baehr, Wolfgang;Frederick, Jeanne M.
通讯作者：
Frederick, Jeanne M.
影响因子：
3.4
作者：
Caminos, Elena;Vaquero, Cecilia F.;Martinez-Galan, Juan R.
通讯作者：
Martinez-Galan, Juan R.
影响因子：
1.8
作者：
Sammons, Joshua D.;Gross, Alecia K.
通讯作者：
Gross, Alecia K.
影响因子：
3.5
作者：
Athanasiou D;Aguila M;Opefi CA;South K;Bellingham J;Bevilacqua D;Munro PM;Kanuga N;Mackenzie FE;Dubis AM;Georgiadis A;Graca AB;Pearson RA;Ali RR;Sakami S;Palczewski K;Sherman MY;Reeves PJ;Cheetham ME
通讯作者：
Cheetham ME