Corneal haze phenotype in Aldh3a1-null mice: In vivo confocal microscopy and tissue imaging mass spectrometry
Corneal haze phenotype in Aldh3a1-null mice: In vivo confocal microscopy and tissue imaging mass spectrometry
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DOI：
10.1016/j.cbi.2016.12.017
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发表时间：
2017-10-01
影响因子：
5.1
通讯作者：
Vasiliou, Vasilis
中科院分区：
文献类型：
--
作者：
Chen, Ying;Jester, James V.;Vasiliou, Vasilis
ALDH3A1是一种角膜晶状体蛋白，通过催化和非催化功能保护眼组织免受紫外线照射。此外，ALDH3A1通过同时调节增殖和分化，在角膜上皮细胞的动态平衡中发挥作用。我们之前已经证明，在C57B6/129sV混合遗传背景下的Aldh3a1基因敲除小鼠会患晶状体白内障。在本研究中，我们评估了培育成C57B/6J同源背景(KO)的Aldh3a1基因敲除小鼠的角膜表型。KO和野生型(WT)角膜的体内共聚焦显微镜检查显示，KO小鼠表现出角膜混浊，表现出明显的角膜基质光散射。这种角膜表型进一步用空间分辨率的成像质谱仪(IMS)来表征，它揭示了基于不同脂质定位的三层结构。在这些初步研究中，与WT小鼠相比，KO小鼠的血脂谱没有差异；然而，KO鼠角膜上皮层的酰基CoA结合蛋白(m/z9966)和组蛋白H4.4(m/z 11308)的蛋白质谱变化被发现增加。这是首次使用IMS来表征角膜组织中的内源性蛋白质和脂类，并从分子水平探索角膜混浊的表型。综上所述，本研究提供了第一个由角膜晶体蛋白丢失引起的细胞诱导的角膜混浊的遗传动物模型，并有力地支持了ALDH3A1对细胞透明度至关重要的观点。最后，IMS代表了一种有价值的新方法来揭示角膜疾病背后的分子变化。(C)爱思唯尔爱尔兰有限公司出版的2016年。
ALDH3A1 is a corneal crystallin that protects ocular tissues from ultraviolet radiation through catalytic and non-catalytic functions. In addition, ALDH3A1 plays a functional role in corneal epithelial homeostasis by simultaneously modulating proliferation and differentiation. We have previously shown that Aldh3a1 knockout mice in a C57B6/129sV mixed genetic background develop lens cataracts. In the current study, we evaluated the corneal phenotype of Aldh3a1 knockout mice bred into a C57B/6J congenic background (KO). In vivo confocal microscopy examination of KO and wild-type (WT) corneas revealed KO mice to exhibit corneal haze, manifesting marked light scattering from corneal stroma. This corneal phenotype was further characterized by Imaging Mass Spectrometry (IMS) with spatial resolution that revealed a trilayer structure based on differential lipid localization. In these preliminary studies, no differences were observed in lipid profiles from KO relative to WT mice; however, changes in protein profiles of acyl-CoA binding protein (m/z 9966) and histone H4.4 (m/z 11308) were found to be increased in the corneal epithelial layer of KO mice. This is the first study to use IMS to characterize endogenous proteins and lipids in corneal tissue and to molecularly explore the corneal haze phenotype. Taken together, the current study presents the first genetic animal model of cellular-induced corneal haze due to the loss of a corneal crystallin, and strongly supports the notion that ALDH3A1 is critical to cellular transparency. Finally, IMS represents a valuable new approach to reveal molecular changes underlying corneal disease. (C) 2016 Published by Elsevier Ireland Ltd.