Mouse Mutagenesis: Phenotype-Driven Neuroscience Screens

该提案的总体目标是建立一个中心，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诱变筛选。我们仔细选择了五个表型筛选：1）昼夜节律，2）恐惧条件反射，3）视觉，4）神经内分泌激素，5）对精神兴奋剂的反应。为了使我们包括筛选，我们已经建立了以下一组标准：* 表型的生物学背景必须是成熟的，并且对神经科学具有重要意义; * 表型类别中突变体的特征已经很好地建立; * 表型筛选必须能够自动化和规模化;* 初步筛选必须每年处理最少一万只老鼠; * 参与筛选和跟进工作的研究人员必须是该领域的顶尖专家。我们的目标是：1.进行大规模、全基因组、表型驱动的ENU诱变筛选，以筛选影响神经系统和行为的5个结构域的隐性突变。2.筛选、分离和表征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情境依赖性和线索性恐惧条件反射的突变。4.利用视网膜电图（ERG）、视觉诱发电位（VEP）和眼底照相三种不同的方法筛选、分离和表征改变视力的突变。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、分离和表征改变小鼠对精神兴奋剂治疗反应的突变。7.通过提供“在线”表型筛选分析的快速访问，作为小鼠突变体的国家资源，以便更大的科学界可以访问小鼠。随着人类基因组计划的进展和更多的人类和小鼠基因的序列被确定，大量基因的功能将无法单独通过序列和表达来预测。表型驱动的诱变筛选为理解这些基因的功能提供了一种重要的方法。

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.