Mouse Model of DBC Dysfunction

DBC 功能障碍小鼠模型

• 批准号: 8324574
• 负责人: RONALD G GREGG
• 金额: $ 19.19万
• 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/8324574
• 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; abstracting; 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

Abstract 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.

抽象的 先天性静止性夜盲症（CSNB）是非进行性夜盲症的临床术语 损害视杆细胞介导的视力的视网膜疾病。 CSNB的完整形式（cCSNB） 是由去极化双极细胞 (DBC) 信号转导和 患者与 NYX、GRM6 或 TRPM1 突变有关。因为一切的流动 视觉输入通过双极细胞从外层视网膜转移到内层视网膜，这一点至关重要 了解 DBC 中的信号转导机制。最近的工作已经定义 这个级联中有几个（但不是全部）参与者。在这个项目中，我们将确定另一个关键 蛋白质。在目标 1 中，我们将鉴定新小鼠的基因和突变 DBC 功能障碍模型，nob5。 nob5 基因位点与所有其他已知基因位点不同 DBC 功能障碍模型，因此其鉴定将添加另一种蛋白质 那些已知对 DBC 功能至关重要的物质。这些研究将使用下一代 测序和定位克隆以定位和克隆负责 nob5 的基因 表型。在目标 2 中，我们将定义与视网膜功能相关的 nob5 表型， 使用视网膜电图和视杆细胞和视锥细胞的全细胞膜片钳记录 DBC 和视锥细胞超极化双极细胞，以及突触的形态 在光感受器和 DBC 之间，使用共焦显微镜和 免疫组织化学。该项目完成后，我们将确定一个新的 正常 DBC 功能所需的蛋白质，可用于筛选 cCSNB 患者。