Analysis of the shocked zebrafish motility mutant

描述(由申请人提供)：应聘者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。拟议的项目为斑马鱼运动突变体Shock的研究提供了一个强有力的新培训组成部分，它补充和扩展了从研究生和博士后培训中获得的技能。有100多种斑马鱼基因突变会影响幼鱼的游泳能力。这些突变中的几个已经被证明与人类肌无力综合征有关。这种受电击的突变是不寻常的，因为运动性缺陷会随着年龄的增长而改善。震惊的鱼最初会瘫痪，但在几天的过程中获得游泳的能力。初步数据表明，过度的电耦合是电击鱼有缺陷的游泳表型的基础。尽管脊椎动物的神经肌肉接头是所有突触中研究得最好的，电耦合是所有未成熟脊椎动物肌肉的一个特征，但肌肉细胞之间的电耦合的意义还没有得到解决。这项拟议的研究包括三个目标：1)探索缝隙连接对野生型和休克鱼类肌肉突触生理的功能影响；2)通过位置克隆和测序的并行方法确定休克表型背后的基因；3)了解休克鱼类最初瘫痪及其随后恢复的机制。这项工作将在Paul Brehm博士的实验室进行，Paul Brehm博士是脊椎动物神经肌肉突触和离子通道功能开发的专家。最近，他的实验室专注于分析斑马鱼的运动性突变，并在很短的时间内取得了很大进展，发现了几个运动性突变的分子基础，包括SOFA马铃薯(乙酰胆碱受体增量亚单位)、RELACK(二氢吡啶受体)和Twitch One(Rapsyn)。此外，这些研究对受体和缝隙同步蛋白在构建突触中的作用有了新的见解。保罗·布雷姆博士的实验室是由五名独立研究人员组成的紧密联系的小组的一部分，他们有不同但相关的兴趣，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。

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.