Mouse Model of DBC Dysfunction

摘要 先天性静止性夜盲(CSNB)是非进行性夜盲的临床术语 损害视杆介导的视力的视网膜疾病。CSNB的完整形式(CCSNB) 是由于去极化双极细胞(DBC)信号转导的缺陷和 患者与NYX、GRM6或TRPM1基因突变有关。因为所有人的流动 视觉输入通过双极细胞从视网膜外部传递到内部，这是至关重要的 了解DBCS的信号转导机制。最近的研究已经定义了 在这一系列事件中，有几个，但不是所有的。在这个项目中，我们将确定另一个关键 蛋白。在目标1中，我们将确定一只新老鼠背后的基因和突变 DBC功能障碍模型，nob5。Nob5基因座与所有已知的基因座不同。 DBC功能障碍的模型及其鉴定将增加另一种蛋白质 那些已知对DBC功能至关重要的基因。这些研究将使用下一代 测序和定位克隆定位和克隆nob5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， 应用视杆细胞和视锥细胞视网膜电描记术和全细胞膜片钳记录 DBCS和锥体超极化双极细胞以及突触的形态 在光感受器和DBCS之间，使用共聚焦显微镜和 免疫组织化学。在这个项目完成时，我们将确定一个新的 正常DBC功能所需的、可用于筛选的蛋白质 慢性阻塞性肺疾病患者。

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.