Brain development phenotyping of IMPC lethal mutant mice

IMPC致死突变小鼠的大脑发育表型

基本信息

批准号：
10428657
负责人：
NOELLE D DWYER
金额：
$ 56.14万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10428657
关键词：
Address Affect Age Apical Apoptosis Axon Basal lamina Biological Biology Body Size Brain Brain Diseases Candidate Disease Gene Cell Polarity Cell division Cells Cellular Structures Cephalic Child Health Communities Complex Cytokinesis Data Databases Daughter Defect Development Developmental Biology Disease Disease model Dissection Embryo Ethylnitrosourea Etiology Excision Gene Expression Genes Genetic Goals Heterozygote Histologic Histology Histopathology Homozygote Human Human Development Image Intellectual functioning disability International Investigation Kinesin Kinetics Knock-out Knockout Mice Knowledge Labyrinth Lead Libraries Medicine Methods Microtubules Mission Mitosis Modeling Morphogenesis Morphology Mus Mutant Strains Mice Mutate Mutation National Institute of Child Health and Human Development Neural Tube Defects Neuroanatomy Neurodevelopmental Disorder Organ Outcome Phenotype Placenta Process Proliferating Prosencephalon Proteins Public Health Publishing Research Research Personnel Resources Role Series Shapes Simple Epithelium Spinal Cord Stains Stretching Structural defect Structure Surface Testing Thick Time Tube Ventricular apical membrane base brain abnormalities brain malformation brain size cohort comparison control daughter cell developmental disease experience forward genetics gastrulation gene discovery gene function genetic approach improved inner ear development insight mouse genetics mouse model mutant nerve stem cell neural plate neurodevelopment neuromechanism novel organ growth response reverse genetics self-renewal stem cell division tool

项目摘要

Abstract: The mechanisms by which neural stem cells build the brain from a simple epithelial tube is a compelling mystery of biology. Mouse genetics has been one of the most powerful tools to discover the genes and processes involved in building a healthy brain, or in neurodevelopmental disorders that affect brain structure or function. The library of targeted mutations created by the International Mouse Phenotyping Consortium (IMPC) provides a highly valuable resource for understanding those genes and processes. Many mutations that cause neurodevelopmental phenotypes are lethal, either due to the brain defect or to pleiotropic functions of the gene in other organs essential for viability. Thus, the bank of lethal mutants at IMPC provides an enriched set of candidate genes required for brain development. However, changes in brain structure or wiring are difficult to detect without the right tools and expertise. We propose to apply our expertise to phenotype selected knockout lines that are likely to have neurodevelopment defects, based on gene expression, function, and/or known mutation in a human developmental disorder. This is in response to an FOA to characterize developmental defects in lethal IMPC mutants. We will combine non-hypothesis driven gross phenotyping with hypothesis-driven analysis of a few selected brain development mutants. To add value and efficiency to this screen, subsets of mutants will also be tested by co-investigators for inner ear development phenotypes, and placenta, gastrulation, or neural tube defects. For the first tier of phenotyping, lines will be tested for age of lethality and phenotyped for gross abnormalities of brain, inner ear, spinal cord, and body development. Tier 2 phenotyping will incorporate histopathology of brain, inner ear, and placenta, since many structural defects in these organs cannot be ascertained without sectioning and staining. We will test for proliferation, layering, and axon tract defects. Tier 3 phenotyping will focus on a small number of mutants with both abnormal brain phenotypes and gene functions in cytokinesis, to address our hypothesis: that different defects in cytokinesis of cortical neural stem cells underlie a variety of brain malformations. We will make use of methods we have established for quantitative analysis of cytokinetic furrowing and abscission defects in developing mouse cortex. In all stages of phenotyping, heterozygotes and homozygotes will be compared to controls quantitatively and with statistical rigor. Data will be shared with IMPC for the benefit of the community. Our team of three investigators has a combined >50 years of expertise working on cellular bases of organ development in the mouse model, and forward and reverse genetics. Through this project , we will discover new mouse models for developmental disorders of the brain, inner ear, and placenta, and will provide important new insights into the mechanisms of neural stem cell divisions and cellular defects underlying brain malformations.

摘要：神经干细胞从简单的上皮管中建立大脑的机制是引人入胜的生物学之谜。鼠标遗传学一直是发现基因的最强大工具之一以及涉及建立健康大脑或影响大脑的神经发育障碍的过程结构或功能。国际鼠标表型创建的目标突变库财团（IMPC）为理解这些基因和过程提供了高度宝贵的资源。许多引起神经发育表型的突变是致命的，要么是由于脑缺陷或多效性造成的基因在其他器官中的功能对于生存力必不可少。因此，IMPC的致命突变体库提供大脑发育所需的一组丰富的候选基因。但是，大脑结构的变化或没有正确的工具和专业知识，很难检测到布线。我们建议将我们的专业知识应用于基于基因的表型选定的敲除线可能具有神经发育缺陷人类发育障碍中的表达，功能和/或已知突变。这是对 FOA表征致命IMPC突变体中发育缺陷。我们将结合非假设驱动的通过对一些选定的脑发育突变体的假设驱动分析进行总表型。增加价值和对此屏幕的效率，突变体的子集也将通过共同研究器对内耳进行测试发育表型和胎盘，胃造成或神经管缺陷。对于表型的第一层，线条将测试杀伤年龄和表型，以确保大脑，内耳，脊髓，脊髓的严重异常和身体发展。第2层表型将结合大脑，内耳和胎盘的组织病理学，由于如果没有切片和染色，就无法确定这些器官中的许多结构缺陷。我们将测试增殖，分层和轴突缺陷。 3级表型将集中在少数具有异常脑表型和基因在细胞因子中的突变体，以解决我们的假设：皮质神经干细胞细胞因子的不同缺陷是多种脑畸形的基础。我们将利用我们已经建立的方法来定量分析细胞动力学的皱纹和脱落开发鼠标皮质的缺陷。在表型的各个阶段，杂合子和纯合子将是与对照定量和统计严格相比。数据将与IMPC共享，以获取社区。我们的三个调查人员团队的组合> 50年的专业知识在蜂窝方面工作小鼠模型中器官发育的基础，并向前和逆转遗传学。通过这个项目，我们将发现新的鼠标模型，以实现大脑，内耳和胎盘的发育障碍，并将提供有关神经干细胞分裂和细胞缺陷机制的重要新见解潜在的大脑畸形。