Alpha Crystallin Function in the Zebrafish

描述(申请人提供)：脊椎动物晶状体蛋白在晶状体发育和维持晶状体透明度方面起着重要作用。？？晶体蛋白的异常表达也与许多人类的眼部、肌肉和神经系统疾病有关，最近还与各种癌症有关。我们在这项提案中的目标是利用斑马鱼作为模型系统的优势来1)详细说明晶状体蛋白在晶状体发育中的作用，以及2)在小鼠和斑马鱼1？-晶状体蛋白启动子中确定新的调控区域。这些研究将有助于我们对正常和病理晶状体功能的理解。我们建议通过使用合成的称为吗啉的反义寡核苷酸分子来单独或联合抑制斑马鱼晶状体蛋白的翻译，来检验晶状体蛋白在斑马鱼和哺乳动物之间保守的作用这一假设。这项工作将为分析？？的特性提供有效的模型系统。晶体蛋白的发育功能，并将使未来的转基因晶体蛋白的体内研究。斑马鱼中存在两个不寻常的1？-晶体蛋白(一个是晶状体特有的，一个是在体内普遍表达的)，这将使我们能够研究哺乳动物1？-晶体蛋白的功能是如何在两种斑马鱼同类之间分配的。斑马鱼也将被用作快速筛选系统，以确定控制1？？-晶体蛋白的时间和空间表达的新的调控区域。到目前为止，调控元件仅在哺乳动物1β-晶体蛋白启动子的相对较小的近端区域已知。未来的研究可以利用这个斑马鱼模型来测试体内突变启动子对晶状体蛋白表达的影响。拟议的工作将在本科生生物系完成，让学生在视觉科学中使用的实验设计、数据分析和尖端研究技术方面获得经验。 与公众健康相关：晶状体蛋白是眼镜片发育和维持晶状体透明度所必需的。这些蛋白质的突变和数量异常与多种眼部和神经系统疾病有关，并在各种癌症中发现。该项目将使用斑马鱼作为模式物种，研究晶状体蛋白在晶状体发育中的作用以及控制其生产的机制。

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.