Mouse Mutagenesis: Phenotype-Driven Neuroscience Screens

这项提议的总体目标是创建一个中心，该中心将专注于与神经系统和行为相关的五个表型领域的大规模ENU突变筛选。我们谨慎地选择了五个表型筛选：1)昼夜节律，2)恐惧条件作用，3)视觉，4)神经内分泌激素，5)对精神刺激的反应。为了纳入筛选，我们制定了以下一套标准：*表型的生物学背景必须成熟，并对神经科学具有重要意义；*表型类突变的特征已经确立；*表型筛选必须能够实现自动化和规模化；*初始筛选必须能够每年至少处理10,000只小鼠；*参与筛选的研究人员及其后续工作必须是该领域的领先专家。我们的目标是：1.进行大规模、全基因组、表型驱动的ENU突变筛选，筛选针对影响神经系统和行为的五个结构域的隐性突变。2.筛选、分离和鉴定改变小鼠昼夜节律表型的突变。3.筛选、分离和表征改变小鼠背景依赖和线索恐惧条件反射的突变。4.利用视网膜电信号(ERG)、视觉诱发电位(VEP)和眼底照相三种不同的方法筛选、分离和鉴定改变视力的突变。5.筛选、分离和鉴定改变下丘脑-肾上腺(HPA)轴和下丘脑-甲状腺(HPT)轴的突变。6.筛选、分离和鉴定改变小鼠对精神刺激药物治疗反应的突变。7.作为小鼠突变体的国家资源，“在线”提供快速的表型筛选分析，以便更广泛的科学界能够接触到小鼠。随着人类基因组计划的进展，以及更多人和鼠基因的序列被确定，大量基因的功能将不能仅通过序列和表达来预测。表型驱动的突变筛选为了解这些基因的功能提供了重要的途径。

The overall objectives of this proposal are to create a Center that will focus on large-scale ENU mutagenesis screens in five phenotypic domains relevant to the nervous system and behavior. We have carefully chosen to focus upon five phenotypic screens: 1) circadian rhythms, 2) fear conditioning, 3) vision, 4) neuroendocrine hormones, and 5) response to psychostimulants. In order for us to include a screen, we have established the following set of criteria: * the biological context of the phenotype must be mature and of significance to neuroscience; * the characterization of mutants in the phenotypic class is well established; * the phenotypic screen must be amenable to automation and scaling; * the initial screen must be capable of a throughput of at least 10,000 mice per year; * the investigators involved in the screens and their follow up must be leading experts in the field. Our aims are: 1. To conduct a large-scale, genome-wide, phenotype-driven ENU mutagenesis screen for recessive mutations that targets five domains influencing the nervous system and behavior. 2. To screen, isolate and characterize mutations that alter the circadian phenotype of mice. 3. To screen, isolate and characterize mutations that alter context- dependent and cued fear conditioning in mice. 4. To screen, isolate and characterize mutations that alter vision using three different methods: electroretinogram (ERG), visually evoked potentials (VEP) and fundus photography. 5. To screen, isolate and characterize mutations that alter the hypothalmic-adrenal (HPA) axis and the hypothalamic-thyroid (HPT) axis. 6. To screen, isolate and characterize mutations that alter the response of mice to psychostimulant treatment. 7. To act as a national resource for mouse mutants by providing rapid access to phenotypic screening analyses "online" so that mice are accessible to the greater scientific community. As the human genome project progresses and the sequences of more human and mouse genes are determined, the function of a large number of genes will not be predictable by sequence and expression alone. Phenotype-driven mutagenesis screens provide an important approach to understand the function of these genes.