Deletion of Seventeen Amino Acids at the C-Terminal End of Aquaporin 0 Causes Distortion Aberration and Cataract in the Lenses of AQP0ΔC/ΔC Mice.
Deletion of Seventeen Amino Acids at the C-Terminal End of Aquaporin 0 Causes Distortion Aberration and Cataract in the Lenses of AQP0ΔC/ΔC Mice.
复制标题
删除水通道蛋白 0 C 末端的 17 个氨基酸会导致 AQP0ΔC/ΔC 小鼠晶状体畸变像差和白内障。
DOI：
10.1167/iovs.18-26378
复制
发表时间：
2019
影响因子：
4.4
通讯作者：
Kumari,Sindhu
中科院分区：
文献类型：
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作者：
Varadaraj,Kulandaiappan;Kumari,Sindhu
目的：方法：建立敲入（knockin，KI）小鼠模型（AQP 0 ΔC/ΔC），在透镜中以末端切割形式表达AQP 0，观察AQP 0缺失对透镜透明性、聚焦性和稳态的影响。为此，将AQP 0基因工程改造为C-末端切割有氨基酸1至246，而不是野生型（WT）的全长1至263。在验证KI整合到基因组中及其表达后，将小鼠模型繁殖几代。在不同发育阶段对AQP 0 KI纯合（AQP 0 ΔC/ΔC）和杂合（AQP 0 +/ΔC）晶状体进行成像和透明度分析。相应地，透镜中的像差使用标准金属网格聚焦方法来表征。将数据与年龄匹配的WT、AQP 0敲除（AQP 0-/-）和AQP 0杂合（AQP 0 +/-）晶状体进行比较。结果：AQP 0 ΔC/ΔC晶状体在整个胚胎发育过程中直至产后第15天（P15）都是透明的。与年龄匹配的AQP 0-/-晶状体相反，AQP 0-/-晶状体在胚胎期本身就会发生白内障。然而，在P5时，AQP 0 ΔC/ΔC透镜存在畸变像差;在P15后，白内障开始发展并且进展速度超过年龄匹配的AQP 0 −/−透镜。与AQP 0 +/-晶状体相比，AQP 0 +/ΔC晶状体在1岁时就已透明，但从P15开始出现畸变像差。结论：完整和端裂的AQP 0在透镜中从外皮质到内核的特定分布是建立折射率梯度以实现无像差的适当聚焦和保持透明度所必需的。
Purpose: Investigate the effects of the absence of 17 amino acids at the C-terminal end of Aquaporin 0 (AQP0) on lens transparency, focusing property, and homeostasis.Methods: A knockin (KI) mouse model (AQP0 ΔC/ΔC) was developed to express AQP0 only as the end-cleaved form in the lens. For this, AQP0 was genetically engineered as C-terminally end-cleaved with amino acids 1 to 246, instead of the full length 1 to 263 of the wild type (WT). After verifying the KI integration into the genome and its expression, the mouse model was bred for several generations. AQP0 KI homozygous (AQP0 ΔC/ΔC) and heterozygous (AQP0+/ΔC) lenses were imaged and analyzed at different developmental stages for transparency. Correspondingly, aberrations in the lens were characterized using the standard metal grid focusing method. Data were compared with age-matched WT, AQP0 knockout (AQP0−/−), and AQP0 heterozygous (AQP0+/−) lenses.Results: AQP0 ΔC/ΔC lenses were transparent throughout the embryonic development and until postnatal day 15 (P15) in contrast to age-matched AQP0−/− lenses, which developed cataract at embryonic stage itself. However, there was distortion aberration in AQP0 ΔC/ΔC lens at P5; after P15, cataract began to develop and progressed faster surpassing that of age-matched AQP0−/− lenses. AQP0+/ΔC lenses were transparent even at the age of 1 year in contrast to AQP0+/− lenses; however, there was distortion aberration starting at P15.Conclusions: A specific distribution profile of intact and end-cleaved AQP0 from the outer cortex to the inner nucleus is required in the lens for establishing refractive index gradient to enable proper focusing without aberrations and for maintaining transparency.