Using the C. elegans Oocyte to Model the Cell Biology of Early Onset Dystonia

使用线虫卵母细胞模拟早发性肌张力障碍的细胞生物学

• 批准号: 9021284
• 负责人: David Irwin Greenstein
• 金额: $ 22.8万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9021284
• 关键词: Address; Age; Animals; Biochemical; Biological; Caenorhabditis elegans; Cell Nucleus; Cell physiology; Cells; Cellular biology; Child; Defect; Development; Dystonia; Early Onset Dystonia; Embryo; Embryonic Development; Essential Tremor; Experimental Models; Genes; Genetic; Genetic Models; Genetic Screening; Glutamic Acid; Goals; Growth; Homologous Gene; Human; Image; Inherited; Life; Limb structure; Mediating; Meiosis; Messenger RNA; Modeling; Molecular; Molecular Genetics; Movement Disorders; Muscle Contraction; Mutation; Nematoda; Neurologic; Neurons; Nuclear Envelope; Nuclear Export; Oocytes; Oogenesis; Parkinson Disease; Pathogenesis; Pathway interactions; Positioning Attribute; Posture; Prevalence; Process; Protein Biosynthesis; Proteins; Recovery of Function; Research; Resolution; Ribonucleoproteins; Role; Site; Suppressor Mutations; Symptoms; Synapses; System; Testing; TorsinA; Translating; Translational Regulation; Walking; Work; base; cellular imaging; early onset; experience; functional restoration; genetic manipulation; high reward; high risk; insight; mRNA Export; mutant; neuromuscular function; novel; particle; protein complex; protein function; restoration; temporal measurement; tool; trafficking

ABSTRACT Early onset dystonia or DYT1 dystonia is a debilitating neurological movement disorder that first presents in children at a mean age of about 12. DYT1 dystonia is caused by a mutation in the DYT1/Tor1a gene that encodes the evolutionarily conserved torsinA protein resulting in the deletion of a single glutamic acid residue (ΔE302/303 or ΔE). The mechanism through which the ΔE mutation causes DYT1 dystonia is unclear because the basic cellular function of torsinA is unknown. Intriguing new work suggests that torsinA might function in a new cellular mechanism by which large ribonucleoprotein particles (RNPs) are exported from the nucleus via budding through the nuclear envelope. When this function of torsinA is perturbed, messenger RNAs appear to inefficiently traffic to their synaptic sites of protein synthesis, compromising neuromuscular function. The field needs to know whether this new model is correct, and if so, to identify the specific molecular mechanisms by which torsinA promotes nuclear envelope budding for RNP export. This application seeks to address these goals using the oocytes of the nematode Caenorhabditis elegans as a tractable experimental model. Mutations in the best characterized C. elegans torsinA homolog (called ooc-5 for oocyte formation abnormal five) cause defects in oocyte growth. The oocyte growth defect is the earliest developmental abnormality observed in ooc-5 mutants and therefore is likely reflective of the primary underlying biochemical and cell biological deficits observed in cells lacking torsinA/OOC-5 function. Our hypothesis is that ooc-5 mutations disrupt oogenesis by interfering in part with the nuclear export assembly, or function of oocyte growth-promoting RNPs that we have defined. Given its extensive evolutionary conservation, OOC-5/torsinA is likely to perform the same elemental protein function in C. elegans oocytes and mammalian neurons. Interestingly, prior work has shown that the mechanisms of translational regulation discovered in C. elegans oocytes are also important for the development and function mammalian neurons. This proposal capitalizes on the many experimental advantages afforded by the C. elegans germline system, including the ability to conduct high-resolution live-cell imaging and the ease of molecular genetic and biochemical manipulations. Further, our group has spent two decades pioneering the mechanisms controlling oocyte development in C. elegans. Recently, we defined proteins and mRNA components of RNPs that regulate the growth, meiotic development, and developmental potential of C. elegans oocytes. Our research team is thus ideally positioned to critically test the generality of an RNP budding role for torsinA. To assess this new role for torsinA, we will: (1) Analyze the cellular mechanisms by which OOC-5 controls the oocyte growth process; and (2) Define genetic networks that can restore function to ooc-5/torsinA mutants.

抽象的 早期发作肌张力障碍或dyt1a是一种令人衰弱的神经运动器滴度，呈现在 平均年龄大约12岁的孩子。肌张力障碍因dyt1/tor1a基因突变而造成caud ud ud 编码进化保守的龙田蛋白 （ΔE302/303或ΔE）。 Torsina的基本细胞功能是未知的。 大型核糖核蛋白颗粒（RNP）通过核从细胞核中导出的新细胞机制 通过核包膜发芽。 无效的蛋白质合成的突触部位，损害了神经肌肉功能。 需要知道这个新模型是否正确，如果是这样 Torsina促进了RNP出口的核包络。 该申请旨在使用线虫Caenorhabditis的卵母细胞解决这些目标 秀丽隐杆线作为一种可牵引的实验模型。 （称为卵母细胞形成异常的OOC-5）导致卵母细胞生长的缺陷。 在OOC-5突变体中观察到的高光发育异常，因此可能反映了您 在缺乏Torsina/OOC-5功能的细胞中观察到的主要潜在生化和细胞生物学缺陷。 我们的假设是OOC-5突变通过部分干扰核的卵子发生 我们定义的组装或促卵母细胞生长的RNP的功能。 保护，OOC-5/Torsina可能在秀丽隐杆线虫卵母细胞中执行相同的元素蛋白功能，并且 哺乳动物神经元。 在秀丽隐杆线虫中发现的卵母细胞对于发育和功能哺乳动物神经元也很重要。 该提案利用了C.秀丽隐杆线虫种系系统提供的许多实验优势， 包括进行高分辨率活细胞成像的能力以及分子遗传和 生化操纵。 秀丽隐杆线虫的卵母细胞发育。 调节秀丽隐杆线虫卵巢的生长，减数分裂的发育和发育潜力 因此，团队是理想的定位，可以严格测试滚动角色的一般性 Torsina的新作用，我们将：（1）分析OOC-5控制卵母细胞生长的细胞机制 过程；（2）定义可以将功能恢复为OOC-5/Torsina muts的遗传网络。