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

• 批准号: 8586073
• 负责人: Brian D Perkins
• 金额: $ 21.91万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8586073
• 关键词: Role; Wrb; Vertebrate; Ribbon; Synapse

Project Summary/Abstract The synapses of vertebrate photoreceptors and sensory hair cells release neurotransmitter through specialized structures known as synaptic ribbons. While ribbon synapses and conventional synapses utilize similar mechanisms for synaptic vesicle release, the protein composition at each type of synapse can vary. In a screen for insertional mutations affecting visual function in zebrafish, we identified the zebrafish pinball wizard 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, reduction in the number 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 retina function through a combination of histological, physiological, and molecular approaches. In Aim 1, we will identify where Wrb blocks synaptic transmission by examining immunohistochemical markers for various synaptic proteins and through transgenic and electrophysiological recordings. 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 ribbon synapses and possibly provide insight into candidate genes for blindness/deafness disorders.

项目总结/摘要 脊椎动物光感受器和感觉毛细胞的突触释放神经递质 通过被称为突触带的特殊结构。而带状突触和 传统的突触利用类似的突触囊泡释放机制， 每种类型的突触的组成可以不同。在一个筛选插入突变影响 在斑马鱼的视觉功能中，我们鉴定了斑马鱼弹球向导突变体， 光感受器退化和听力损失。突变破坏了wrb基因， 编码一种新的170个氨基酸的蛋白质，其功能未知。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， 光感受器突触处停靠的条带数量，并显著降低ERG 应答这些数据有力地表明，BMPb可能是完整地参与带状突触 组装和/或功能。为了验证这一假设，我们将研究斑马鱼中的BMPB的作用 视网膜功能通过组织学、生理学和分子方法的组合。 在目标1中，我们将通过检查 免疫组织化学标记的各种突触蛋白，并通过转基因和 电生理记录。在目标2中，我们将使用一个 转基因方法纯化带标签的Bismb复合物和随后的质量分析 谱这些研究将为深入了解BMPB的作用提供帮助，BMPB是一种新的蛋白质， 是带状突触结构和功能的关键组成部分。binding的定义 合作伙伴和随后的表征，在体内系统中的BCLB将打开新的 研究带状突触的途径，并可能提供深入了解候选基因， 失明/耳聋疾病。