Characterization and Genetic Analysis of Basal Ganglia Axon Pathfinding

描述（由申请人提供）：基底神经节的发育及其连接的形成是一个复杂且鲜为人知的过程。基底神经节是运动功能、认知和语言发展以及社会和情绪调节的组成部分。虽然一些基因已被确定在基底节区神经元规范或基底节区疾病中起作用，但这些基因和其他基因如何调节基底节区连通性的发展尚不清楚。此外，发育过程中基底神经节轴突寻路的实际过程尚未被表征。我的假设是纹状体和中间脑多巴胺能（mesDA）神经元的连接是由机器人、狭缝和ephslephrins的寻路基因家族控制的。我的长期目标是表征基底节区连通性的发育和遗传控制，使用斑马鱼（Danio rerio）作为模型系统。为了探索基底神经节连接的发展，本项目有三个具体目的：目的1。开发分子和遗传标记可视化纹状体和中间脑多巴胺能（mesDA）神经元及其轴突。我正在使用原位基因标记物和抗体来标记这些细胞核。为了研究它们的寻路，我已经生成了新的基底节区神经元特异性增强子系，包括dlx(mini):gfp和foxP2-enhancerA:egfp。目标2。表征斑马鱼纹状体和mesDA神经元在发育过程中的正常轴突寻路。我将使用增强子：gfp构建体和转基因系来描述连通性的发展，并将寻路与比率、缝隙、ephs和ephrin的表达模式进行比较。目标3。评估robo、slit和ephphrin基因在纹状体和mesDA神经元寻路中的作用。我将通过使用突变鱼线和morpholinos的组合来测试这些基因家族中不同成员的功能和影响。临床意义：我们的结果将提高我们对基底神经节连通性发展的理解，这在神经发育和神经退行性疾病中受到影响。摘要：本文所描述的工作将是对基底神经节轴突寻路的首次描述，描述了连通性的发展及其遗传基础。我的研究结果将包括对基底神经节发育的描述，以及对特定基因作用的分析。这个项目包括一个结构良好的职业发展计划，广泛的机构资源支持和国际知名的导师，以帮助我在K奖期间过渡到独立的终身教职岗位。

DESCRIPTION (provided by applicant): The development of the basal ganglia and the formation of their connections are complex and poorly understood processes. The basal ganglia are integral to motor function, to cognitive and language development, and to social and emotional regulation. While some genes have been identified with roles in basal ganglia neuron specification, or in diseases of the basal ganglia, how these and other genes regulate the development of basal ganglia connectivity is not known. Further, the actual process of basal ganglia axon pathfinding during development has not been characterized. My hypothesis is that connectivity of the striatum and the mesodiencephalic dopaminergic (mesDA) neurons is controlled by the pathfinding gene families of the robos, the slits, and the ephslephrins. My long-term objective is to characterize the development and genetic control of basal ganglia connectivity, using zebrafish (Danio rerio) as a model system. In order to explore the development of basal ganglia connections, this project has three specific aims: Aim 1. Develop molecular and genetic markers to visualize striatal and mesodiencephalic dopaminergic (mesDA) neurons and their axons. I am using in situ gene markers and antibodies to label these nuclei. To study their pathfinding, I have generated novel enhancer lines specific for basal ganglia neurons, including dlx(mini):gfp and foxP2-enhancerA:egfp. Aim 2. Characterize the normal axon pathfinding of the zebrafish striatum and mesDA neurons during development. I will use enhancer:gfp constructs and transgenic lines to describe the development of connectivity, and compare the pathfinding to the expression patterns of the ratios, slits, ephs, and ephrins. Aim 3. Evaluate the role of robo, slit, and ephlephrin genes in striatal and mesDA neuron pathfinding. I will test the function and effects of different members of these gene families by using a combination of mutant fish lines and morpholinos. Clinical Significance: Our results will improve our understanding of the development of basal ganglia connectivity, which is affected in neurodevelopmental and neurodegenerative disorders. Summary: The work described here will be the first description of axon pathfinding of the basal ganglia, characterizing both the development of connectivity and its genetic basis. My results will include both descriptive elements of basal ganglia development, as well as analyses of the role of specific genes. This project consists of a well-structured career development plan, extensively supported with institutional resources and an internationally known mentor, to assist me with the transition during the K award to an independent tenure-track faculty position.