权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The Role of Wrb in Vertebrate Ribbon Synapse Formation

Wrb 在脊椎动物带状突触形成中的作用

基本信息

批准号：
8489300
负责人：
Brian D Perkins
金额：
$ 18.64万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8489300
关键词：
Affect Afferent Neurons Affinity Chromatography Amino Acids Anatomy Antibodies Architecture Auditory Binding Biochemistry Biological Assay Blindness Candidate Disease Gene Cells Coiled-Coil Domain Communication Complex Data Defect Development Disease Docking Ear Electron Microscopy Endocytosis Exhibits Functional disorder Gene Expression Genes Hair Cells Hearing Human Immunohistochemistry In Situ Hybridization Investigation Knowledge Labyrinth Lead Learning Mass Spectrum Analysis Membrane Membrane Potentials Modeling Molecular Molecular Probes Morphology Mutate Mutation Neurons Perception Phenotype Photoreceptors Property Protein Binding Proteins Reagent Research Retina Role Sensory Sensory Hair Stimulus Structure Synapses Synaptic Cleft Synaptic Transmission Synaptic Vesicles System Testing Transgenes Transgenic Animals Transgenic Organisms Transmembrane Domain Vertebrate Photoreceptors Vesicle Vision Visual Zebrafish abstracting base deafness hearing impairment in vivo insight interdisciplinary approach mutant neurotransmitter release novel photoreceptor degeneration postsynaptic presynaptic protein protein interaction research study response ribbon synapse synaptic function synaptogenesis

项目摘要

Project Summary/Abstract The synapses of vertebrate photoreceptors and sensory hair cells release neurotransmitter through specialized structures known as synaptic ribbons. In a screen for insertional mutations affecting visual function in zebrafish, we identified the zebrafish wrb mutant, which exhibited photoreceptor degeneration and hearing loss. The mutation disrupts the wrb gene, which encodes a novel 170 amino acid protein with no known function. In preliminary studies, we found that wrb mutants exhibited mislocalization of ribbon synapse components, reduced numbers of docked ribbons at photoreceptor synapses, and significantly reduced ERG responses. These data strongly suggest that Wrb could be integrally involved in ribbon synapse assembly and/or function. To validate this hypothesis, we will study the role of Wrb in zebrafish synaptic function through a combination of histological and molecular approaches. In Aim 1, we will identify the subcellular localization of Wrb using transgenic approaches, we will determine if Wrb affects ribbon assembly during development, how loss of Wrb affects dendritic morphology of postsynaptic cells, and whether Wrb blocks synaptic vesicle release or membrane endocytosis. In Aim 2, we will identify binding partners of Wrb using a transgenic approach to purify tagged Wrb complexes and subsequent analysis by mass spectroscopy. These studies will provide insights into the role of Wrb, a novel protein that may be a critical component to ribbon synapse structure and function. The identity of Wrb binding partners and the subsequent characterization of Wrb in an in vivo system will open new avenues of research on synaptic architecture and possibly provide insight into candidate genes for blindness/deafness disorders.

项目概要/摘要脊椎动物感光器和感觉毛细胞的突触释放神经递质通过称为突触带的特殊结构。插入突变筛选影响斑马鱼视觉功能的，我们鉴定了斑马鱼wrb突变体，它表现出光感受器退化和听力损失。该突变破坏了wrb基因，编码一种新的 170 个氨基酸的蛋白质，其功能尚不清楚。在初步研究中，我们发现wrb突变体表现出带状突触成分的错误定位，减少了光感受器突触处的对接丝带数量，并且 ERG 显着降低回应。这些数据强烈表明 Wrb 可能整体参与带状突触组装和/或功能。为了验证这一假设，我们将研究 Wrb 在斑马鱼中的作用通过组织学和分子方法相结合的突触功能。在目标 1 中，我们将使用转基因方法鉴定 Wrb 的亚细胞定位，我们将确定是否 Wrb 在发育过程中影响带状组装，Wrb 的损失如何影响树突形态突触后细胞的数量，以及 Wrb 是否阻断突触小泡释放或膜内吞作用。在目标 2 中，我们将使用转基因方法鉴定 Wrb 的结合伙伴纯化标记的 Wrb 复合物并通过质谱进行后续分析。这些研究将深入了解 Wrb 的作用，Wrb 是一种新型蛋白质，可能是带状突触的结构和功能。 Wrb结合伙伴的身份和后续 Wrb 在体内系统中的表征将为突触研究开辟新途径结构并可能提供对失明/耳聋疾病候选基因的深入了解。