Analysis of neurodevelopment using mouse mutagenesis

使用小鼠诱变分析神经发育

• 批准号: 7631348
• 负责人: DAVID R. BEIER
• 金额: $ 47.07万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7631348
• 关键词: Adult; Affect; American; Arts; Autistic Disorder; Brain; Brain Diseases; Brain Neoplasms; Chemicals; Chromosome Mapping; Cloning; Congenital Abnormality; DNA; Defect; Degenerative Disorder; Development; Development Plans; Disease; Embryonic Development; Epilepsy; Genes; Genetic; Genome; Goals; Hand; Histology; Human; Huntington Disease; In Situ; Institutes; Lead; Learning Disabilities; Life; Mental Retardation; Metabolic Diseases; Methods; Modeling; Mus; Mutagenesis; Mutant Strains Mice; Mutate; Mutation; Nature; Neurons; Nitrosourea Compounds; Organ; Parkinson Disease; Pathway interactions; Pattern; Phenotype; Plant Roots; Prosencephalon; RNA Interference; Reporter Genes; Research Personnel; Schizophrenia; Science; Screening procedure; Staging; Structure; Syndrome; Validation; Work; developmental disease; high throughput technology; insight; interest; malformation; migration; mutant; neurodevelopment; positional cloning; public health relevance; rapid technique

DESCRIPTION (provided by applicant): Mouse mutants that perturb cortical patterning have provided considerable insight into the development of the mammalian brain. We have previously undertaken a project to generate models of human congenital defects by screening ENU-mutagenized mice for recessive mutations affecting late embryonic development. The screen incorporated a genetic mapping component, with the intent to facilitate the positional cloning and functional characterization of the mutant genes. The strategy has worked well, and we have generated many mice with phenotypes similar to human malformation syndromes and birth defects. In this proposal we aim to target cortical development, with the goal of identifying and cloning additional mutants that will be useful for understanding how the mammalian brain is patterned. Specifically, we plan to screen for mice that have abnormalities of brain morpholology and histology. We will also screen for mutations that perturb patterning of reporter genes that mark specific anatomical structures, A third strategy is to screen for mutations that have a genetic interaction with known cortical patterning genes such as Lis1. We have developed efficient strategies for genetic mapping and positional cloning, and we are currently working on rapid methods of mutation validation using RNAi. Finally, for a subset of mutations that have relatively specific effects on cortical development, we plan to pursue in-depth analysis, including a collaborative effort with investigators at the Allen Institute for Brain Science utilizing high-throughput technologies for in situ expression analysis. Depending upon the nature of the mutated gene, we will pursue functional studies as appropriate. PUBLIC HEALTH RELEVANCE. We propose to treat mice with the chemical ethyl-nitrosourea (ENU), which causes mutations in DNA. We will examine the progeny of treated mice to assess whether they have disorders of brain development. We are particularly interested in those that have relatively subtle effects; that is, which appear to specifically affect the formation of the brain, but not other organs. Once we have generated these mutants, we will use state-of-the-art methods of genome analysis to identify the mutated gene. With these in hand, we can begin to evaluate what developmental pathways are affected in these abnormal mice. Understanding the formation of the brain will likely help us to understand the roots of the many brain diseases affecting one in five Americans today (www.ninds.nih.gov). These diseases include developmental disorders (including autism), degenerative diseases of adult life, metabolic diseases, and brain tumors. Defects in neuronal migration within the forebrain lead to mental retardation, epilepsy and severe learning disabilities. The forebrain is also primarily affected in many other diseases such as Parkinson's and Huntington's diseases and schizophrenia. Significant efforts, both public and private, are currently underway to begin to identify more of the genes expressed in the brain at various stages as well as their expression patterns. This information, combined with the mutational analysis we plan, will help uncover the basic steps required for normal brain development.

描述（由申请人提供）：扰动皮质图案的小鼠突变体对哺乳动物大脑的发育提供了相当大的见解。 我们以前已经进行了一个项目，以通过筛选核糖核酸小鼠来产生人类先天性缺陷的模型，以实现影响晚期胚胎发育的隐性突变。 屏幕融合了遗传学映射成分，目的是促进突变基因的位置克隆和功能表征。 该策略效果很好，我们已经产生了许多具有类似人类畸形综合症和先天缺陷的表型的小鼠。 在此提案中，我们旨在瞄准皮质发育，目的是确定和克隆其他突变体，这些突变体对了解哺乳动物大脑的图案方式有用。 具体而言，我们计划筛选具有脑形态学和组织学异常异常的小鼠。 我们还将筛选出突变的突变，这些突变扰乱了标记特定解剖结构的报告基因的模式，第三种策略是筛选与已知皮质模式基因（如Lis1）具有遗传相互作用的突变。 我们已经开发了有效的遗传图和位置克隆策略，目前我们正在使用RNAi进行快速的突变验证方法。 最后，对于对皮质发育具有相对特定影响的突变子集，我们计划进行深入分析，包括与艾伦脑科学研究所的研究人员的合作努力，利用高通量技术来进行现场表达分析。 根据突变基因的性质，我们将适当进行功能研究。 公共卫生相关性。 我们建议用化学乙烯乙酸（ENU）治疗小鼠，该化学乙基环（ENU）在DNA中引起突变。 我们将检查治疗小鼠的后代，以评估它们是否患有大脑发育障碍。 我们对那些具有相对微妙的影响的人特别感兴趣。也就是说，这似乎特别影响大脑的形成，而不是其他器官。 一旦我们产生了这些突变体，我们将使用最新的基因组分析方法来识别突变的基因。 借助这些，我们可以开始评估这些异常小鼠受影响的发育途径。 了解大脑的形成可能会帮助我们了解影响当今五分之一美国人的许多​​脑部疾病的根源（www.ninds.nih.gov）。 这些疾病包括发育障碍（包括自闭症），成人生活的退化性疾病，代谢疾病和脑肿瘤。 前脑内神经元迁移的缺陷导致智力低下，癫痫和严重的学习障碍。 前脑也主要在许多其他疾病中受到影响，例如帕金森氏症和亨廷顿的疾病和精神分裂症。 目前正在进行的重要努力，无论是公共还是私人的努力，都开始在各个阶段及其表达模式中确定更多在大脑中表达的基因。 这些信息与我们计划的突变分析相结合，将有助于发现正常大脑发育所需的基本步骤。