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）：视网膜的感光细胞和双极细胞以及听觉和前庭系统的毛细胞通过神经递质释放的分级变化发出感觉刺激信号。为此，这些细胞进化出了突触带，即束缚大量细胞的蛋白质结构。突触带功能的分子机制目前尚不清楚，少数蛋白质定位于突触带和突触带。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剥离。 Ribeye 是由转录辅助抑制子 CtBP2 的另一个起始位点形成的。Ribeye 的确切作用仍然未知，该提案的长期目标是确定其功能作用。为了研究 Ribeye 的功能，我们将结合分子生物学、遗传学和电生理学，主要使用斑马鱼作为主要模型系统，我们生成并表征了两个 ribeye Aim 基因的靶向突变。 2，我们研究一种或两种 Ribeye 基因产物对光感受器和毛细胞带结构的影响 在目标 3 中，我们将研究 Ribeye 去除对分布的影响。在目标 4 中，我们将研究 Ribeye 损失对神经丘毛细胞胞吐作用和钙电流的影响，从而在分子水平上了解突触带功能将最终有助于理解视觉和听觉信息的处理方式。此外，它可能提供帮助理解特别影响视觉和听力的疾病的线索。此外，对这些特殊神经元突触前过程的基本了解将对一般神经元交流产生更广泛的影响。可能有助于我们了解心理健康和神经系统疾病的各个方面。