Alpha Crystallin Function in the Zebrafish

斑马鱼的α晶状体蛋白功能

• 批准号: 7780221
• 负责人: Mason Posner
• 金额: $ 18.45万
• 依托单位: ASHLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7780221
• 关键词: Affect; Antisense Oligonucleotides; Automobile Driving; Biological Assay; Biological Models; Biology; Cataract; Cell Differentiation process; Cells; Crystalline Lens; Crystallins; Data; Data Analyses; Development; Disease; Effectiveness; Embryo; Experimental Designs; Eye; Future; Genetically Engineered Mouse; Goals; Green Fluorescent Proteins; Heat shock proteins; Histology; Homeostasis; Lead; Lens Fiber; Maintenance; Malignant Neoplasms; Mammals; Modeling; Mus; Muscle; Mutate; Mutation; Nervous system structure; Neuro-Ocular System; Neurological observations; Nucleic Acid Regulatory Sequences; Oligonucleotides; Physiological; Play; Production; Promoter Regions; Proteins; Regulatory Element; Reporter Genes; Research Technics; Role; Screening procedure; Staging; Students; System; Systems Analysis; Techniques; Technology; Testing; Time; Translations; Two-Dimensional Gel Electrophoresis; Western Blotting; Work; Zebrafish; alpha-Crystallins; experience; fiber cell; genetic manipulation; high throughput screening; human disease; in utero; in vivo; lens; lens transparency; mouse development; novel; paralogous gene; promoter; protein expression; protein function; public health relevance; research study; tool; vision science

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.

描述（申请人提供）：脊椎动物？？晶状体蛋白在透镜发育和保持透镜透明度中起重要作用。异常的表情？晶体蛋白还涉及许多人类的眼睛、肌肉和神经系统疾病，并且最近与各种癌症有关。我们在这个提案中的目标是利用斑马鱼作为模型系统的好处，1）详细说明？？晶状体蛋白在透镜发育中的作用; 2）在小鼠和斑马鱼中鉴定新的调节区域。- 晶体蛋白启动子。这些研究将有助于我们了解正常和病理透镜功能。我们建议测试的假设，作用？？透镜发育中的晶状体蛋白在斑马鱼和哺乳动物之间是保守的，通过使用称为morpholinos的合成反义寡核苷酸分子来抑制每个斑马鱼的翻译。晶体蛋白单独和组合。这项工作将提供一个有效的模型系统分析的具体？？晶体蛋白发育功能，并将允许未来的体内研究转基因？？晶体蛋白。两个不寻常的存在1？？- 斑马鱼中的晶状体蛋白（一种透镜特异性，另一种在体内普遍表达）将使我们能够研究哺乳动物1？晶体蛋白已经在两种斑马鱼旁系同源物之间分开。斑马鱼也将被用来作为一个快速筛选系统，以确定新的调控区域控制1？- 晶体蛋白的时空表达。调控元件是迄今为止只知道从一个相对较小的近端区域的哺乳动物1？？- 晶体蛋白启动子。未来的研究可以利用这种斑马鱼模型来测试突变的启动子对？晶状体蛋白表达。拟议的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。 公共卫生相关性：？？晶状体蛋白是眼透镜发育和保持透镜透明度所必需的。这些蛋白质的突变和异常数量与多种眼睛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型物种来研究？？晶体蛋白在透镜发育中的作用及其产生机制。