Models for Vision Research

项目总结/摘要 全世界约有5000万人失明，约1.5亿人视力严重受损。 除了外伤和感染外，大多数人类眼病都是遗传性的。的数量 引起视网膜疾病的人类基因座比可获得的相关动物模型的数量大约9倍， 这表明在研究已知发生在人类身上的疾病的模型方面存在很大的差距。老鼠和它的 发达的遗传学，与人类生理学和解剖学的相似性，以及遗传学的可及性， 操纵是被广泛接受的和有用的模型系统。小鼠模型已被用于提供 用于人类疾病的候选基因，用于在整个发育和疾病进展中研究的组织，以及 用于治疗的测试系统。它们也是一个理想的平台，以确定和解剖生物相关的 通过遗传途径。 在上一个融资周期中，我们生成了超过60个具有眼部缺陷的模型。我们已经鉴定出 基于20个突变株系，从中获得了许多独特的见解。在这个应用程序中，我们计划 完成剩余40个品系的分子和表型表征（目标1）， 提供给科学研究界。 扩展我们正在进行的遗传研究，我们建议使用敏化化学诱变筛选， 揭示Crumbs 1途径中的重要途径（目的2）。虽然有许多策略可供选择， 鉴定主要基因/突变的相互作用因子，化学诱导突变具有的优点是， 它们将允许对与CRUMBS 1相互作用的大量基因进行无偏鉴定。这些 基因可以解释与Crumbs 1内突变相关的大量疾病。它还将允许 鉴定与CRUMBS 1胞外结构域相互作用的因子， 目前可用的方法难以处理。在本申请中，我们将筛选约10，000个诱变的 G3 Crumbs 1 rd 8/rd 8小鼠通过间接检眼镜检查鉴定存在改变的Crumbs 1 rd 8的突变体 眼底表型这些因素的分子基础将被确定，并通过使用标准 免疫组织化学方法结合使用4Pi显微镜，我们将检查的影响， CRUMBS 1通路上新发现的基因/突变。 该方案的成功完成不仅将产生良好表征的眼部模型，而且将 可能识别CRUMBS 1通路的入口点以及其他重要的分子， 眼睛生物学，并为我们提供了建立和测试有关正常眼功能的假设的机会， 疾病病理学

PROJECT SUMMARY/ABSTRACT Approximately 50 million people worldwide are blind and ~150 million are significantly vision impaired. Except for trauma and infections, the majority of human eye diseases are genetic in nature. The number of human loci causing retinal disease is ~ 9-fold greater than the number of available associated animal models, indicating a large gap in models for studying diseases that are known to occur in humans. The mouse with its well-developed genetics, similarity to human physiology and anatomy, and accessibility for genetic manipulation is a widely accepted and useful model system. Mouse models have been used to provide candidate genes for human diseases, tissues for study throughout development and disease progression, and test systems for therapies. They are also an ideal platform to identify and dissect biologically relevant pathways through genetic means. In the last funding cycle, we generated >60 models with ocular defects. We have identified the molecular basis of 20 of the mutant lines from which many unique insights were obtained. In this application, we plan to complete the molecular and phenotypic characterization of the 40 remaining lines (Aim 1) and make them available to the scientific research community. Extending our ongoing genetic studies, we propose to use a sensitized chemical mutagenesis screen to reveal pathways important in the Crumbs1 pathway (Aim 2). While there are many strategies available to identify interacting factors of primary genes/mutations, chemically induced mutations have the advantage that they will allow for the unbiased identification of a wide array of genes that interact with CRUMBS1. These genes may explain the plethora of diseases associated with mutations within Crumbs1. It will also allow for identification of factors that interact with the extracellular domain of CRUMBS1, an endeavor that has been intractable by the current available methods. In the present application, we will screen ~10,000 mutagenized G3 Crumbs1rd8/rd8 mice by indirect ophthalmoscopy to identify mutants that present with an altered Crumbs1drd8 fundus phenotype. The molecular bases of these factors will be identified and through the use of standard immunohistochemical methodologies in conjunction with the use of 4Pi microscopy, we will examine the effects of the newly identified genes/mutations on the CRUMBS1 pathway. Successful conclusion of this proposal will not only generate well characterized ocular models, but will potentially identify entry points into the CRUMBS1 pathways as well as other molecules that are important in eye biology and afford us the opportunity to build and test hypotheses about normal ocular function and disease pathology.