Rpe65-/- and Lrat-/- mice:: Comparable models of Leber congenital amaurosis
Rpe65-/- and Lrat-/- mice:: Comparable models of Leber congenital amaurosis
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DOI：
10.1167/iovs.08-1727
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发表时间：
2008-06-01
影响因子：
4.4
通讯作者：
Crouch, Rosalie K.
中科院分区：
文献类型：
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作者：
Fan, Jie;Rohrer, Baerbel;Crouch, Rosalie K.
目的。用作莱伯先天性黑蒙模型的 Rpe65(-/-) 小鼠具有缓慢的视杆细胞退化和快速的视锥细胞丧失，可能是由于视锥细胞视蛋白的错误运输所致。该动物不产生11-顺式视网膜，并且通过施用11-顺式视网膜可恢复视锥细胞损失和视杆细胞反应。同样，Lrat(-/-) 小鼠不产生 11-顺式视网膜。作者试图确定 Lrat(-/-) 小鼠中是否也存在对 Rpe65(-/-) 小鼠视杆细胞和视锥细胞视蛋白的相同影响，从而确定这些变化可归因于 11-cis 视网膜的缺乏，而不是 RPE65 的某些未知功能。通过免疫组织化学方法定位视杆细胞和视锥细胞视蛋白。通过用 11-顺式视网膜再生来测定功能性视蛋白水平。异视紫红质水平通过色素提取来测定。通过质谱法测定视蛋白磷酸化。结果。两种模型中的视杆细胞均缓慢退化。在研究的 6 个月时间过程中，可再生视杆细胞视蛋白水平相似，视杆细胞视蛋白以低水平磷酸化（约 10%），并且通过非光过程产生最小的 9-顺式视网膜，产生微量光响应。在这两个模型中，S-视蛋白和 M/L-视蛋白未能适当地运输到视锥细胞外节，并且发生了快速的视锥细胞退化。两种模型中视锥细胞视蛋白的错误运输在 11-cis 视网膜给药后均被阻止。结论。这些数据表明，Lrat(-/-) 和 Rpe65(-/-) 小鼠是莱伯先天性黑蒙研究的可比模型，并且破坏性视锥细胞视蛋白错误运输是由 11-cis 视网膜缺乏引起的。
PURPOSE. The Rpe65(-/-) mouse, used as a model for Leber congenital amaurosis, has slow rod degeneration and rapid cone loss, presumably because of the mistrafficking of cone opsins. This animal does not generate 11-cis retinal, and both cone loss and rod response are restored by 11-cis retinal administration. Similarly, the Lrat(-/-) mouse does not produce 11-cis retinal. The authors sought to determine whether the same effects on rod and cone opsins in the Rpe65(-/-) mouse are also present in the Lrat(-/-) mouse, thereby establishing that these changes can be attributed to the lack of 11-cis retinal rather than to some unknown function of RPE65.METHODS. Rod and cone opsins were localized by immunohistochemical methods. Functional opsin levels were determined by regeneration with 11-cis retinal. Isorhodopsin levels were determined from pigment extraction. Opsin phosphorylation was determined by mass spectrometry.RESULTS. Rods in both models degenerated slowly. Regenerable rod opsin levels were similar over the 6-month time course investigated, rod opsin was phosphorylated at a low level (approximately 10%), and minimal 9-cis retinal was generated by a nonphotic process, giving a trace light response. In both models, S-opsin and M/L-opsin failed to traffic to the cone outer segments appropriately, and rapid cone degeneration occurred. Cone opsin mistrafficking in both models was arrested on 11-cis retinal administration.CONCLUSIONS. These data show that the Lrat(-/-) and Rpe65(-/-) mice are comparable models for studies of Leber congenital amaurosis and that the destructive cone opsin mistrafficking is caused by the lack of 11-cis retinal.