Analysis of the "shocked" zebrafish motility mutant

“震惊”的斑马鱼运动突变体分析

• 批准号: 6757752
• 负责人: Julia Eve Dallman
• 金额: $ 11.73万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6757752
• 关键词: biotechnology; chordate locomotion; developmental genetics; diving /swimming; electromyography; electrophysiology; functional ability; gap junctions; gene mutation; genetic mapping; genetic markers; membrane channels; molecular cloning; neuromuscular disorder; paralysis; polymerase chain reaction; protein sequence; receptor coupling; receptor expression; synaptogenesis; video microscopy; voltage /patch clamp; western blottings; wild animals; zebrafish

DESCRIPTION (provided by applicant): The applicant's goal is to obtain a position as Assistant Professor at an academic institution. The conventional route to this goal has been thwarted by childbirth, followed by a debilitating series of treatments for cancer. The proposed project provides a strong component of new training for the study of a zebrafish motility mutant, shocked, which compliments and extends skills acquired from graduate student and post-doctoral training. Over one hundred zebrafish mutations exist that affect the ability of a young fish to swim. Several of these mutations have already proved relevant to human myasthenic syndromes. The shocked mutation is unusual in that the motility defect improves with age. Shocked fish are initially paralyzed but acquire the ability to swim over the course of several days. Preliminary data suggests that excessive electrical coupling underlies the defective swimming phenotype in shocked fish. Although the vertebrate neuromuscular junction is the best studied of any synapse and electrical coupling is a feature of all immature vertebrate muscle, the significance of electrical coupling among muscle cells has not been addressed. The proposed research includes three aims: 1) To pursue the functional consequences of gap junctions for muscle synaptic physiology in wild type and shocked fish; 2) to identify the gene that underlies the shocked phenotype by parallel approaches of positional cloning and sequencing likely candidates; and 3) to understand the mechanisms that underlie the initial paralysis in shocked fish as well as their subsequent recovery. This work will be carried out in the laboratory of Dr. Paul Brehm, an expert in development of the vertebrate neuromuscular synapse and ion channel function. More recently his laboratory has focused on analyzing zebrafish motility mutants and has made great progress in a short time, discovering the molecular basis for several motility mutants including sofa potato (acetylcholine receptor delta subunit), relaxed (dihydropyridine receptor), and twitch once (rapsyn). Moreover, these studies have yielded new insight into the roles of the receptor and rapsyn in structuring the synapse. Dr. Paul Brehm's laboratory is part of a tight knit group of five independent researchers with distinct but related interests and shared microscopy and molecular facilities that create an environment with ample resources to carry out the proposed research.

描述（由申请人提供）：申请人的目标是在学术机构获得助理教授的职位。实现这一目标的传统途径受到了分娩的阻碍，随后是一系列令人衰弱的癌症治疗。拟议的项目提供了一个强大的组成部分，新的培训研究的斑马鱼运动突变，震惊，恭维和扩展从研究生和博士后培训获得的技能。超过100种斑马鱼突变影响了幼鱼的游泳能力。这些突变中的一些已经被证明与人类肌无力综合征有关。休克突变是不寻常的，因为运动缺陷随着年龄的增长而改善。休克的鱼最初会瘫痪，但在几天内获得游泳的能力。初步数据表明，过度的电耦合的基础上有缺陷的游泳表型休克鱼。虽然脊椎动物神经肌肉接头是所有突触中研究最好的，并且电耦合是所有未成熟脊椎动物肌肉的特征，但肌细胞之间电耦合的意义尚未得到解决。拟议的研究包括三个目标：1）追求野生型和休克鱼的肌肉突触生理学的间隙连接的功能后果; 2）通过位置克隆和测序可能的候选人的平行方法来确定休克表型的基因;和3）了解休克鱼最初瘫痪以及随后恢复的机制。这项工作将在脊椎动物神经肌肉突触和离子通道功能开发专家Paul Bengym博士的实验室进行。最近，他的实验室专注于分析斑马鱼运动突变体，并在短时间内取得了很大进展，发现了几种运动突变体的分子基础，包括沙发土豆（乙酰胆碱受体δ亚基），松弛（二氢吡啶受体）和抽搐一次（rapsyn）。此外，这些研究对受体和rapsyn在构建突触中的作用产生了新的见解。Paul Bavelm博士的实验室是一个由五名独立研究人员组成的紧密组织的一部分，这些研究人员具有不同但相关的兴趣，并共享显微镜和分子设施，为开展拟议的研究创造了充足的资源环境。