Mutation in the Zebrafish cct2 Gene Leads to Abnormalities of Cell Cycle and Cell Death in the Retina: A Model of CCT2-Related Leber Congenital Amaurosis.
Mutation in the Zebrafish cct2 Gene Leads to Abnormalities of Cell Cycle and Cell Death in the Retina: A Model of CCT2-Related Leber Congenital Amaurosis.
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DOI：
10.1167/iovs.17-22919
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发表时间：
2018-02-01
影响因子：
4.4
通讯作者：
Tomarev SI
中科院分区：
文献类型：
--
作者：
Minegishi Y;Nakaya N;Tomarev SI
包含TCP-1的伴侣蛋白β亚基（CCT）由CCT 2编码的复合杂合突变导致Leber先天性黑蒙（LCA）。在这项研究中，一个cct 2突变系的斑马鱼，研究CCT 2突变的作用，在脊椎动物的LCA。使用CRISPR-Cas9系统产生Cct 2突变体斑马鱼系。发展中的野生型和突变幼虫的眼睛的变化进行了监测，使用显微镜，免疫染色，TUNEL，和EdU测定。通过将CCT 2 RNA注射到斑马鱼胚胎中来研究突变表型的表型拯救。cct 2突变（L394 H-7 del）导致合成了一种突变的cctβ蛋白，其中L394 H置换并缺失7个氨基酸残基（位置395-401）。纯合的cct 2-L394 H-7 del突变体在受精后2天（dpf）表现出小眼表型，并且在5 dpf后胚胎致死。与野生型相比，纯合cct 2-L394 H-7 del突变体在2dpf时视网膜神经节细胞分化减弱，视网膜细胞周期受到影响，神经视网膜细胞死亡显著增加。注射编码野生型人CCTβ的RNA挽救了小眼表型，减少了视网膜细胞死亡，并恢复了CCTβ蛋白和主要客户蛋白Gβ1的水平，与野生型相比，这些蛋白在纯合cct 2-L394 H-7 del突变体中显著降低。这些结果表明cct 2通过调节细胞周期在视网膜发育中起着重要作用。在纯合cct 2-L394 H-7 del突变体中观察到的视网膜病理学类似于人LCA患者的视网膜病理学。
The compound heterozygous mutations in the β subunit of chaperonin containing TCP-1 (CCT), encoded by CCT2, lead to the Leber congenital amaurosis (LCA). In this study, a cct2 mutant line of zebrafish was established to investigate the role of CCT2 mutations in LCA in vertebrates. A cct2 mutant zebrafish line was produced using the CRISPR-Cas9 system. Changes in the eyes of developing wild-type and mutant larvae were monitored using microscopy, immunostaining, TUNEL, and EdU assays. Phenotypic rescue of mutant phenotype was investigated by injection of CCT2 RNA into zebrafish embryos. The cct2 mutation (L394H-7del) led to the synthesis of a mutated cctβ protein with the L394H replacement and deletion of 7 amino acid residues (positions 395–401). The homozygous cct2-L394H-7del mutant exhibited a small eye phenotype at 2 days post fertilization (dpf) and was embryonically lethal after 5 dpf. In homozygous cct2-L394H-7del mutants, the retinal ganglion cell differentiation was attenuated, retinal cell cycle was affected, and the neural retinal cell death was significantly increased at 2 dpf compared with wild-type. Injection of RNA encoding wild-type human CCTβ rescued the small eye phenotype, reduced retinal cell death, and restored the levels of CCTβ protein and the major client protein Gβ1 that were significantly reduced in the homozygous cct2-L394H-7del mutant compared with wild-type. These results indicate that cct2 plays an essential role in retinal development by regulating the cell cycle. The retinal pathology observed in the homozygous cct2-L394H-7del mutants resembles the retinal pathology of human LCA patients.
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DOI：
10.1073/pnas.1222663110
发表时间：
2013-02-19
影响因子：
11.1
作者：
Sontag, Emily M.;Joachimiak, Lukasz A.;Thompson, Leslie M.
通讯作者：
Thompson, Leslie M.
影响因子：
9
作者：
Lin, Y-F;Lee, Y-F;Liang, P-H
通讯作者：
Liang, P-H
CCT 复合物通过自噬限制神经病原蛋白聚集。
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
3.4
作者：
Edwards, Malia M.;McLeod, D. Scott;Li, Renzhong;Grebe, Rhonda;Bhutto, Imran;Mu, Xiuqian;Lutty, Gerard A.
通讯作者：
Lutty, Gerard A.
影响因子：
64.5
作者：
Ditzel, L;Löwe, J;Steinbacher, S
通讯作者：
Steinbacher, S