Investigating the Role of Ribeye in Retinal Ribbon Function

研究 Ribeye 在视网膜功能中的作用

• 批准号: 8963532
• 负责人: DAVID Paul ZENISEK
• 金额: $ 41.63万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8963532
• 关键词: Affect; Animals; Auditory; Binding; Binding Sites; Biological Models; Calcium; Cells; Cleaved cell; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; DNA Sequence Alteration; Defect; Dimerization; Disease; Effectiveness; Electron Microscopy; Electrons; Electrophysiology (science); Excision; Exocytosis; Follow-Up Studies; Gene Expression; Generations; Genes; Genetic; Goals; Grant; Hair Cells; Hearing; Individual; Labyrinth; Left; Mental Health; Microscopy; Missense Mutation; Molecular; Molecular Biology; Mutation; Neurons; Oligonucleotides; Phenotype; Photoreceptors; Process; Proteins; Retina; Retinal; Role; Scaffolding Protein; Sensory; Shapes; Signal Transduction; Site; Structure; Synapses; Synaptic Vesicles; System; Techniques; Testing; Transcription Repressor/Corepressor; Transgenic Animals; Vision; Visual; Zebrafish; Zinc Fingers; genome editing; immunoreactivity; insight; knockout animal; lateral line; mutant; nervous system disorder; neuromast; neurotransmitter release; nuclease; presynaptic; protein distribution; public health relevance; ribbon synapse; sensory stimulus; voltage clamp

 DESCRIPTION (provided by applicant): Photoreceptors and bipolar cells of the retina and hair cells of the auditory and vestibular systems signal sensory stimuli as graded changes in neurotransmitter release. To do so, these cells have evolved synaptic ribbons, proteinaceous structures that tether large numbers of synaptic vesicles near release sites. The molecular machinery underlying synaptic ribbon function is poorly understood. A handful of proteins have been localized to the synaptic ribbon and the importance of these individual molecules as well as how they contribute to the unique functions of the synaptic ribbon remains elusive. Of these proteins, the most abundant is Ribeye, a protein unique to the synaptic ribbon and thought to constitute most of the synaptic ribbon and hypothesized to form the core of the synaptic ribbon. Ribeye arises from an alternative start site of the transcriptional corepressor CtBP2. The precise role of Ribeye remains unknown and the long-term goal of this proposal is to determine the functional role of Ribeye in the synaptic ribbon. To study Ribeye function, we will employ a combination of molecular biology, genetic and electrophysiology primarily using zebrafish as a primary model system. In Aim 1 we generated and characterize zebrafish with targeted mutations in both ribeye genes. In Aim 2, we the effect of one or both Ribeye gene products on the structure of photoreceptor and hair cell ribbons. In Aim 3, we will look at the effect of Ribey removal on the distribution and localization of other synaptic ribbon proteins. In Aim 4, we will investigate the effects of Ribeye loss on exocytosis and calcium current in neuromast hair cells. Understanding synaptic ribbon function at the molecular level will ultimately aid in understanding how visual and auditory information is processed and communicated. In addition, it may provide clues to help understand diseases that specifically affect vision and hearing. In addition, the fundamental understanding of presynaptic processes in these specialized neurons will have broader implications for neuronal communication in general and thus, may contribute to our understanding of various aspects of mental health and neurological disorders.

 描述（由申请人提供）：视网膜的光感受器和双极细胞以及听觉和前庭系统的毛细胞将感觉刺激信号作为神经递质释放的分级变化。为了做到这一点，这些细胞已经进化出突触带，这是一种蛋白质结构，将大量突触囊泡拴在释放部位附近。突触带功能的分子机制还知之甚少。少数蛋白质已经定位于突触带，这些单个分子的重要性以及它们如何对突触带的独特功能做出贡献仍然难以捉摸。在这些蛋白质中，最丰富的是Ribeye，一种突触带特有的蛋白质，被认为构成了突触带的大部分，并被假设形成了突触带的核心。Ribeye来自转录辅阻遏物CtBP2的替代起始位点。Ribeye的确切作用仍然未知，该提案的长期目标是确定Ribeye在突触带中的功能作用。为了研究Ribeye功能，我们将采用分子生物学，遗传学和电生理学的组合，主要使用斑马鱼作为主要模型系统。在目标1中，我们产生并表征了在两个ribeye基因中具有靶向突变的斑马鱼。目的二是研究Ribeye基因产物对毛细胞和感光细胞结构的影响。在目标3中，我们将研究Ribey去除对其他突触带蛋白的分布和定位的影响。在目的4中，我们将研究Ribeye损失对神经肥大毛细胞胞吐和钙电流的影响。在分子水平上理解突触带的功能将最终有助于理解视觉和听觉信息是如何处理和交流的。此外，它还可以提供线索，帮助了解专门影响视力和听力的疾病。此外，对这些特化神经元中突触前过程的基本理解将对神经元通信产生更广泛的影响，因此可能有助于我们对精神健康和神经系统疾病的各个方面的理解。