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）：申请人的目标是获得学术机构助理教授的职位。传统的通往这一目标的途径受到分娩的挫败，随后是一系列令人衰弱的癌症治疗方法。拟议的项目为研究斑马鱼运动突变体的研究提供了强大的新培训组成部分，该突变体令人震惊，该突变体震惊，该突变体称赞并扩展了从研究生和博士后培训中获得的技能。存在一百多个斑马鱼突变，影响了年轻鱼游泳的能力。这些突变中的一些已经证明与人肌综合症有关。令人震惊的突变是不寻常的，因为运动缺陷随着年龄的增长而改善。震惊的鱼最初是瘫痪的，但可以在几天内游泳的能力。初步数据表明，过度的电耦合是冲击鱼中有缺陷的游泳表型的基础。尽管脊椎动物神经肌肉连接是任何突触的最佳研究，并且电耦合是所有未成熟脊椎动物肌肉的特征，但尚未解决肌肉细胞中电耦合的重要性。拟议的研究包括三个目的：1）追求野生型和震惊的鱼类肌肉突触生理学间隙的功能后果； 2）确定通过位置克隆和测序可能候选者的平行方法来识别震惊表型的基因； 3）了解震惊鱼类及其随后恢复的最初瘫痪的机制。这项工作将在Paul Brehm博士的实验室中进行，Paul Brehm博士是脊椎动物神经肌肉突触和离子通道功能的专家。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长足的进步，发现了几种运动性突变体的分子基础，包括沙发（乙酰胆碱受体Delta subunit），放松（Dihydropyrpyridine受体）和Twitch（Rapsyn）。此外，这些研究已经对受体和Rapsyn在构建突触中的作用产生了新的见解。保罗·布雷姆（Paul Brehm）博士的实验室是一个由五个独立研究人员组成的紧密编织组的一部分，这些研究人员具有独特但相关的兴趣以及共享的显微镜和分子设施，这些设施创造了一个具有充足资源的环境，以进行拟议的研究。