Cilia Assembly and Transport in Photoreceptor Cells

感光细胞中纤毛的组装和运输

基本信息

批准号：
8723212
负责人：
Brian D Perkins
金额：
$ 44.21万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8723212
关键词：
Address Adult Alleles Amino Acids Anatomy Apical Bardet-Biedl Syndrome Binding Biological Models Blindness Cell Polarity Cells Cilia Clinical Complex Data Defect Disease Docking Dynein ATPase Electron Microscopy Electrons Embryo Gene Components Gene Expression Gene Mutation Genes Genetic Goals Guanosine Triphosphate Phosphohydrolases Human Individual Joubert syndrome Kidney Diseases Lead Link Location Maintenance Mental Retardation Microscopic Microtubules Modeling Molecular Mutation Organelles Pathology Pathway interactions Pattern Phenotype Photoreceptors Polydactyly Population Positioning Attribute Process Proteins Reagent Regulation Retina Retinal Retinal Degeneration Retinal Diseases Retinal Dystrophy Role Side Signal Pathway Signal Transduction Site Situs Inversus Stereotyping Structure Surface System TNFRSF5 gene Techniques Testing Tissues Transgenic Organisms Vertebrate Photoreceptors Vertebrates Zebrafish base ciliopathy cilium biogenesis dynactin hereditary blindness kinetosome light microscopy loss of function migration mutant novel null mutation photoreceptor degeneration protein transport research study retinal damage therapy development tool trafficking

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to understand the molecular basis of cilia formation and maintenance in vertebrate photoreceptor cells. In vertebrates, the assembly and maintenance of photoreceptor outer segments begins with the formation of a connecting cilium. The connecting cilium contains a microtubule- based axoneme that is anchored to the apical inner segment by a basal body. Cilia formation begins with the docking of basal bodies at the apical surface of the inner segment. Genetic mutations disrupting the assembly, structure, or function of basal bodies and/or cilia result in a spectrum of diseases known as ciliopathies. These multisyndromic disorders often present with retinal degeneration, kidney disease, mental retardation, and polydactyly. In the current application, we will utilize loss-of-function strategies in zebrafish to investigate the mechanisms controlling basal body localization. In Specific Aim 1, we will test the hypothesis that the dynein/dynactin complex regulates the apical transport of basal bodies preceding cilia formation by examining zebrafish mutants in dynein and the p150 and p50 subunits of dynactin. In Specific Aim 2, we provide preliminary evidence that basal bodies show a highly polarized arrangement within the adult zebrafish retina. We will directly test the hypothesis that the PCP pathway regulates this patterning and is essential for photoreceptor survival. In Specific Aim 3, we will examine zebrafish carrying null mutations in the Joubert Syndrome gene arl13b for retinal phenotypes. Proposed experiments will also test the requirement of the GTPase domain and a ciliary-targeting sequence RVxPx for Arl13b function. We will also test arl13b for functional interactions with Bardet-Biedl Syndrome (BBS) genes, and components of the PCP pathway. These interactions will identify potential second-site modifiers that enhance expression of photoreceptor phenotypes. The results of these studies will reveal novel mechanisms required for basal body placement prior to cilia formation and to identify novel genetic interactions that influence hereditary blindness.

描述（由申请人提供）：该项目的长期目标是了解脊椎动物感光细胞纤毛形成和维持的分子基础。在脊椎动物中，感光器外节的组装和维护始于连接纤毛的形成。连接纤毛包含基于微管的轴丝，通过基体锚定到顶端内段。纤毛的形成始于基体在内节顶端表面的对接。破坏基体和/或纤毛的组装、结构或功能的基因突变导致一系列称为纤毛病的疾病。这些多综合征通常表现为视网膜变性、肾脏疾病、智力低下和多指畸形。在当前的应用中，我们将利用斑马鱼的功能丧失策略来研究控制基底体定位的机制。在具体目标 1 中，我们将通过检查动力蛋白以及动力蛋白 p150 和 p50 亚基中的斑马鱼突变体来检验动力蛋白/动力蛋白复合物在纤毛形成之前调节基体顶端运输的假设。在具体目标 2 中，我们提供了初步证据，表明成年斑马鱼视网膜内的基底体显示出高度极化的排列。我们将直接检验 PCP 通路调节这种模式并且对于光感受器存活至关重要的假设。在具体目标 3 中，我们将检查 Joubert 综合征基因 arl13b 携带无效突变的斑马鱼的视网膜表型。拟议的实验还将测试 Arl13b 功能对 GTPase 结构域和纤毛靶向序列 RVxPx 的要求。我们还将测试 arl13b 与 Bardet-Biedl 综合征 (BBS) 基因和 PCP 途径组件的功能相互作用。这些相互作用将识别潜在的第二位点修饰剂，增强光感受器表型的表达。这些研究的结果将揭示纤毛形成之前基底体放置所需的新机制，并确定影响遗传性失明的新遗传相互作用。