Alpha Crystallin Function in the Zebrafish

描述（由申请人提供）：脊椎动物？？晶体蛋白在晶状体发育和晶状体透明度的维持中起着至关重要的作用。异常表达？？晶体蛋白还与人类眼睛、肌肉和神经系统的许多疾病有关，最近还与各种癌症有关。我们在这个提案中的目标是利用斑马鱼作为一个模型系统的好处来详细说明？？晶状体发育中的晶体蛋白和2)在小鼠和斑马鱼中发现新的调控区域1？晶状体蛋白启动子。这些研究将有助于我们对正常和病理晶状体功能的理解。我们建议验证？？晶状体发育中的晶状体蛋白在斑马鱼和哺乳动物之间是保守的，这是通过合成的反义寡核苷酸分子来抑制斑马鱼晶状体的翻译。单独或组合结晶。这项工作将为分析？？晶体蛋白的发育功能，并将允许未来转基因晶体蛋白的体内研究。两个“1”的不寻常存在？-斑马鱼中的晶体蛋白（一种晶状体特异性的，另一种在体内普遍表达）将使我们能够研究哺乳动物的功能如何-晶体蛋白在两种斑马鱼的相似物之间已经分化。斑马鱼还将被用作快速筛选系统，以识别控制1?？-晶体蛋白的时空表达。到目前为止，调控元件只在哺乳动物的一个相对较小的近端区域被发现。晶状体蛋白启动子。未来的研究可以利用这个斑马鱼模型来测试突变启动子对？？晶状体蛋白表达。这项工作将在本科生物系完成，让学生获得实验设计，数据分析和视觉科学中使用的前沿研究技术的经验。

DESCRIPTION (provided by applicant): The vertebrate ??crystallins play an essential role in lens development and the maintenance of lens transparency. Abnormal expression of ??crystallins is also involved in numerous human diseases of the eye, muscular and nervous systems, and has recently been implicated in various cancers. Our goals in this proposal are to leverage the benefits of the zebrafish as a model system to 1) detail the roles of ??crystallins in lens development and 2) identify novel regulatory regions in the mouse and zebrafish 1?? -crystallin promoter. These studies will contribute to our understanding of normal and pathological lens function. We propose testing the hypothesis that the role of ??crystallins in lens development is conserved between zebrafish and mammals by using synthetic anti-sense oligonucleotide molecules called morpholinos to inhibit translation of each zebrafish ??crystallin individually and in combination. This work will provide an efficient model system for analyzing the specifics of ?? crystallin developmental function, and will allow future in vivo studies of genetically modified ??crystallins. The unusual presence of two 1?? -crystallins in zebrafish (one lens specific and one ubiquitously expressed in the body) will allow us to examine how the functions of mammalian 1?-crystallin have become divided between the two zebrafish paralogs. The zebrafish will also be used as a rapid screening system to identify novel regulatory regions controlling 1?? -crystallin's temporal and spatial expression. Regulatory elements are so far only known from a relatively small proximal region of the mammalian 1?? -crystallin promoter. Future studies can utilize this zebrafish model to test the in vivo effects of mutated promoters on ??crystallin expression. The proposed work will be done in an undergraduate biology department, allowing students to gain experience in experimental design, data analysis and cutting edge research techniques used in visual science. PUBLIC HEALTH RELEVANCE: ??crystallin proteins are necessary for eye lens development and the maintenance of lens transparency. Mutations in these proteins and abnormal amounts are associated with multiple diseases of the eye and nervous system and found in various cancers. This project will use the zebrafish as a model species for examining the role of ??crystallins in lens development and the mechanisms that control their production.