Mouse Model of DBC Dysfunction

DBC 功能障碍小鼠模型

• 批准号: 8177871
• 负责人: RONALD G GREGG
• 金额: $ 24.26万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8177871
• 关键词: Agonist; Candidate Disease Gene; Cations; Cell physiology; Cells; Cellular Morphology; Cellular Structures; Chromosome Mapping; Clinical; Confocal Microscopy; Defect; Electroretinography; Functional disorder; Gene Mutation; Genes; Glutamate Receptor; Glutamates; Human; Immunohistochemistry; Link; Maps; Mediating; Modeling; Morphology; Mus; Mutant Strains Mice; Mutation; Night Blindness; Patients; Phenotype; Photoreceptors; Proteins; Protocols documentation; Retina; Retinal; Retinal Cone; Retinal Diseases; Role; Signal Pathway; Signal Transduction; Synapses; Techniques; Technology; Testing; Vertebrate Photoreceptors; Vision; Visual; Work; base; gene cloning; human disease; mouse model; mutant; next generation; novel; patch clamp; positional cloning; postsynaptic; protein S precursor; response; retinal rods; ribbon synapse; transmission process

DESCRIPTION (provided by applicant): Congenital stationary night blindness (CSNB) is the clinical term for non-progressive retinal disorders that impair rod-mediated vision. The complete form of CSNB (cCSNB) is caused by defects in depolarizing bipolar cell (DBC) signal transduction and in patients has been linked to mutations in NYX, GRM6 or TRPM1. Because the flow of all visual input is transferred from the outer to the inner retina via bipolar cells, it is critical to understand the mechanism of signal transduction in DBCs. Recent work has defined several, but not all, players in this cascade. In this project, we will identify another key protein. In Aim 1, we will identify the gene and mutation that underlies a new mouse model of DBC dysfunction, nob5. The nob5 gene locus is distinct from all other known models of DBC dysfunction and therefore its identification will add another protein to those known to be critical for DBC function. These studies will use next generation sequencing and positional cloning to map and clone the gene responsible for the nob5 phenotype. In Aim 2, we will define the nob5 phenotype with respect to retinal function, using electroretinography and whole-cell patch clamp recordings from rod and cone DBCs and cone hyperpolarizing bipolar cells, and the morphology of the synapses between photoreceptors and DBCs, using confocal microscopy and immunohistochemistry. At the completion of this project, we will have identified a new protein that is required for normal DBC function and which can be used to screen patients with cCSNB. PUBLIC HEALTH RELEVANCE: This project will make detailed studies of a new mouse model (nob5) that lacks depolarizing bipolar cell (DBC) function. The nob5 gene has not been identified, but is known to involve a protein that has not been previously implicated in DBC function. Identification of the nob5 gene will expand our understanding of DBC signal transduction and will identify a potential new candidate gene for the complete form of human congenital stationary night blindness.

描述（由申请人提供）：先天性静止性夜盲症（CSNB）是一种损害杆介导视力的非进行性视网膜疾病的临床术语。CSNB （cCSNB）的完整形式是由去极化双极细胞（DBC）信号转导缺陷引起的，在患者中与NYX、GRM6或TRPM1突变有关。由于所有视觉输入的流动都是通过双极细胞从外部视网膜转移到内部视网膜，因此了解DBCs的信号转导机制至关重要。最近的研究定义了这个级联中的几个参与者，但不是全部。在这个项目中，我们将确定另一个关键蛋白。在目的1中，我们将鉴定一种新的DBC功能障碍小鼠模型nob5的基因和突变。nob5基因位点不同于所有其他已知的DBC功能障碍模型，因此它的鉴定将为已知的DBC功能关键蛋白增加另一种蛋白质。这些研究将使用下一代测序和定位克隆来定位和克隆nob5表型的基因。在Aim 2中，我们将使用视网膜电图和全细胞膜片钳记录杆状和锥状DBCs以及锥状超极化双极细胞来定义与视网膜功能相关的nob5表型，并使用共聚焦显微镜和免疫组织化学来定义光感受器和DBCs之间的突触形态。在本项目完成后，我们将鉴定出正常DBC功能所需的新蛋白，该蛋白可用于筛查cCSNB患者。