Genetic Screen for Zebrafish Neural Defect Mutants

斑马鱼神经缺陷突变体的遗传筛查

• 批准号: 8117003
• 负责人: Bruce H Appel
• 金额: $ 30.93万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8117003
• 关键词: Affect; Axon; Biological Assay; Biological Models; Cell Communication; Cells; Cellular Morphology; Chemicals; Coupled; Defect; Development; Disease; Embryo; Family; Foundations; Genes; Genetic; Genetic Screening; Goals; Health; Image; Induced Mutation; Injury; Investigation; Knowledge; Larva; Lead; Life; Location; Maps; Mediating; Membrane; Modeling; Molecular; Motor; Mutagenesis; Mutate; Mutation; Nerve; Nervous System Physiology; Nervous system structure; Neuraxis; Neuroglia; Neurons; Oligodendroglia; Peripheral; Peripheral Nerves; Peripheral Nervous System; Phenotype; Population; Proteins; Reporter; Research Personnel; Resolution; Schwann Cells; Series; Stem cells; Techniques; Time; Transgenic Organisms; Work; Zebrafish; base; cell behavior; design; effective therapy; experience; gene discovery; glial cell development; in vivo; injured; migration; mutant; nervous system disorder; neural circuit; neurodevelopment; neuron development; red fluorescent protein; relating to nervous system; repaired; sensory system; transmission process

DESCRIPTION (provided by applicant): The long-term goal of this project is to identify the genes that are required to build a nervous system. The nervous system is composed of two general classes of cells: neurons, which transmit information in the form of electrical impulses, and glial cells, which facilitate impulse transmission and survival of neurons. How neurons and glial cells are formed and organized into functional neural circuits during development is still poorly understood. The project design draws from the well-validated idea that mutations that disrupt neural development reveal critical genes. Building on the expertise of the collaborating team of investigators, this work will discover genes that are necessary for development of motor axons and glial cells that wrap central and peripheral nerves. The project will use zebrafish as a model system, which permits the careful observations necessary to find rare mutations that cause specific neural defects. The first aim of the project is to screen families of mutagenized zebrafish embryos for defects of motor axons and two distinct populations of central and peripheral myelinating glial cells by examining expression of transgenically encoded fluorescent reporter proteins that reveal cell morphologies. The second aim is to determine how each mutation causes its corresponding defect through careful characterization of cellular defects, using immunohistochemical and transgenic cell marking techniques and in vivo time-lapse imaging. The third aim is to determine the chromosomal location of each mutation and identify the affected genes. Completion of this project will enhance understanding of the genetic bases of neurological disorders involving the sensory system or glia and provide genes that might be used to promote neural repair following injury or disease. PUBLIC HEALTH RELEVANCE: This project will identify genes that are necessary to build a nervous system. It will provide a better understanding of how the nervous system develops and potentially lead to new strategies to promote neural repair following disease or injury.

描述(由申请者提供)：该项目的长期目标是确定建立神经系统所需的基因。神经系统一般由两类细胞组成：神经元以电脉冲的形式传递信息，神经胶质细胞促进脉冲的传递和神经元的生存。神经元和神经胶质细胞是如何在发育过程中形成和组织成功能神经回路的，目前还知之甚少。该项目的设计借鉴了一个经过充分验证的想法，即破坏神经发育的突变揭示了关键基因。在合作研究团队的专业知识基础上，这项工作将发现包裹中枢和外周神经的运动轴突和神经胶质细胞发育所必需的基因。该项目将使用斑马鱼作为模型系统，允许进行必要的仔细观察，以发现导致特定神经缺陷的罕见突变。该项目的第一个目标是通过检测揭示细胞形态的转基因编码的荧光报告蛋白的表达，筛选突变斑马鱼胚胎的运动轴突缺陷和两个不同群体的中央和外周髓鞘神经胶质细胞。第二个目标是通过使用免疫组织化学和转基因细胞标记技术以及体内时间推移成像，仔细描述细胞缺陷，确定每个突变是如何导致其相应的缺陷的。第三个目标是确定每个突变的染色体位置，并识别受影响的基因。该项目的完成将加强对涉及感觉系统或神经胶质细胞的神经疾病的遗传基础的了解，并提供可能用于促进损伤或疾病后的神经修复的基因。与公共健康相关：该项目将确定建立神经系统所必需的基因。它将提供更好的了解神经系统是如何发育的，并可能导致促进疾病或损伤后神经修复的新策略。