Genetic Screen for Zebrafish Neural Defect Mutants

描述（由申请人提供）：该项目的长期目标是确定建立神经系统所需的基因。神经系统由两大类细胞组成：神经元，其以电脉冲的形式传输信息，以及神经胶质细胞，其促进脉冲传输和神经元的存活。神经元和神经胶质细胞在发育过程中如何形成和组织成功能性神经回路仍然知之甚少。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。基于研究人员合作团队的专业知识，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，它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。该项目的第一个目的是通过检测转基因编码的荧光报告蛋白的表达来筛选运动轴突缺陷的诱变斑马鱼胚胎家族和两个不同的中枢和外周髓鞘神经胶质细胞群体，这些荧光报告蛋白揭示了细胞形态。第二个目的是通过仔细表征细胞缺陷，使用免疫组织化学和转基因细胞标记技术和体内延时成像，确定每个突变如何导致其相应的缺陷。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。该项目的完成将提高对涉及感觉系统或神经胶质的神经系统疾病的遗传基础的理解，并提供可能用于促进损伤或疾病后神经修复的基因。公共卫生相关性：该项目将确定建立神经系统所必需的基因。它将更好地了解神经系统如何发育，并可能导致新的策略，以促进疾病或损伤后的神经修复。

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.