Models for Vision Research

描述(申请人提供)：全球约有5000万人失明，约1.5亿人视力严重受损。除了创伤和感染，大多数人类眼部疾病本质上都是遗传的。导致视网膜疾病的人类基因座的数量大约是现有相关动物模型数量的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差距。小鼠由于其发达的遗传学、与人类生理和解剖学的相似性以及遗传操作的可及性，是一种被广泛接受和有用的模型系统。小鼠模型已被用于提供人类疾病的候选基因，用于在整个发育和疾病进展过程中进行研究的组织，以及用于治疗的测试系统。它们也是通过遗传手段识别和剖析生物相关途径的理想平台。虽然有许多策略可以用来建立小鼠模型，但化学诱变是制造患有眼病的小鼠最有效的方法之一。Enu诱导的突变允许无偏见地识别对眼睛生物学有不同影响的广泛的基因。在本应用中，我们将通过间接检眼镜、裂隙灯生物显微镜和视网膜电描记术对大约10,000只老年G3小鼠进行筛查。我们的目标是确定50个可遗传的前段或后段异常的模型。重要的是，我们将绘制或补充测试大约30个视网膜异常模型，并使用标准和辅助生殖方法的组合进行位置候选测试或克隆其中的10-20个。具有已证实的可遗传突变的小鼠将通过这笔赠款中建立的网站和研究群体产生的小鼠的完善的JAX分发系统提供给视力研究社区。将在两个适当的年龄为所有模型提供眼前段或眼底照片记录、视网膜电描记术和组织学检查，以确定疾病是静止的还是进展的。在确定了潜在分子基础的视网膜模型中，将通过实时荧光聚合酶链式反应建立发育表达谱，并在表达最高的年龄进行眼睛的原位分析。 这一提议的成功结论不仅将产生具有良好特征性的眼睛模型，还将潜在地确定进入眼睛生物学中重要的新途径的切入点，并为我们提供机会建立和测试关于正常眼功能和疾病病理学的假说。

DESCRIPTION (provided by applicant): Approximately 50 million people world-wide are blind and approximately 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 approximately 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. While there are many strategies available to generate mouse models, chemical mutagenesis is one of the most efficient methods to produce mice with eye diseases. ENU-induced mutations allow unbiased identification of a wide array of genes with different effects on ocular biology. In the present application, we will screen approximately 10,000 aged G3 mice by indirect ophthalmoscopy, slit lamp biomicroscopy, and electroretinography. Our goal is to identify 50 models with heritable anterior segment or posterior segment abnormalities. Importantly, of these, we will map or complementation test approximately 30 models with retinal abnormalities and go on to positionally candidate test or clone 10-20 of these using a combination of standard and assisted reproductive approaches. Mice with proven heritable mutations will be made available to the vision research community through a website established in this grant and through the well-developed JAX distribution system for mice generated from research colonies. Anterior segment or fundus photo-documentation, electroretinography and histology will be provided for all models at two appropriate ages to determine if the disease is stationary or progressive. In the retinal models where the underlying molecular basis is determined, a developmental expression profile will be established by real time PCR, and in situ analysis of the eye will be carried out at an age in which expression is highest. Successful conclusion of this proposal will not only generate well characterized ocular models, but will potentially identify entry points into new pathways that are important in eye biology and afford us the opportunity to build and test hypotheses about normal ocular function and disease pathology.