Analysis of the shocked zebrafish motility mutant

电击斑马鱼运动突变体的分析

基本信息

批准号：
7236025
负责人：
Julia Eve Dallman
金额：
$ 3.81万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2007-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7236025
关键词：
Acetylcholine Address Affect Age Biological Assay Cells Childbirth Chromosome Pairing Class Cloning Connexin 43 Coupling Data Defect Development Dihydropyridine Receptors Electrical Synapse Environment Fishes Gap Junctions Genes Genetic Polymorphism Goals Human Institution Ion Channel Laboratories Link Measures Microscopy Molecular Muscle Muscle Cells Mutate Mutation Myasthenic Syndrome Nature Neuromuscular Junction Paralysed Phenotype Physiological Physiology Positioning Attribute Potato Recovery Research Research Personnel Resources Role Route Series Shock Speed Staging Structure Students Swimming Synapses Techniques Testing Time Training Video Microscopy Work Zebrafish base cancer therapy cell motility day delta opioid receptor extracellular improved inhibitor/antagonist insight interest mutant patch clamp peripheral membrane protein 43K positional cloning post-doctoral training professor receptor skills zebrafish genome

项目摘要

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因分娩以及随后一系列令人衰弱的癌症治疗而受阻。拟议的项目为斑马鱼运动突变体的研究提供了新的培训的重要组成部分，震惊，它补充并扩展了从研究生和博士后培训中获得的技能。斑马鱼存在一百多种突变，这些突变会影响幼鱼的游泳能力。其中一些突变已被证明与人类肌无力综合征有关。这种休克突变很不寻常，因为运动缺陷会随着年龄的增长而改善。受到惊吓的鱼最初会瘫痪，但在几天内就获得了游泳的能力。初步数据表明，过度的电耦合是电击鱼游泳表型缺陷的基础。尽管脊椎动物神经肌肉接头是所有突触中研究得最好的，并且电耦合是所有未成熟脊椎动物肌肉的一个特征，但肌肉细胞之间电耦合的重要性尚未得到解决。拟议的研究包括三个目标：1）探究间隙连接对野生型和电击鱼肌肉突触生理学的功能影响； 2) 通过位置克隆和可能候选基因测序的并行方法来鉴定休克表型背后的基因； 3）了解电击鱼最初瘫痪及其随后恢复的机制。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方面的专家 Paul Brehm 博士的实验室中进行。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巨大进展，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 Brehm 博士的实验室是由五名独立研究人员组成的紧密小组的一部分，他们有着不同但相关的兴趣，并共享显微镜和分子设施，为开展拟议的研究创造了一个拥有充足资源的环境。