Characterization and Genetic Analysis of Basal Ganglia Axon Pathfinding

描述(申请人提供)：基底节的发育及其连接的形成是复杂且知之甚少的过程。基底节对运动功能、认知和语言发育以及社会和情绪调节都是不可或缺的。虽然一些基因已经被确定在基底节神经元规范中的作用，或者在基底节疾病中的作用，但这些基因和其他基因如何调控基底节连通性的发展尚不清楚。此外，基底节轴突在发育过程中的实际过程还没有被描述。我的假设是纹状体和中间脑多巴胺能神经元(MesDA)的连接是由机器人、缝隙和麻黄素等寻路基因家族控制的。我的长期目标是以斑马鱼(Danio Rerio)为模型系统，描述基底节连接的发育和遗传控制。为了探索基底神经节连接的发育，本项目有三个特定的目标：目的1.开发分子和遗传标记以显示纹状体和中间脑的多巴胺能神经元及其轴突。我正在使用原位基因标记和抗体来标记这些细胞核。为了研究他们的路径发现，我已经产生了新的针对基底节神经元的增强子系，包括DLX(Mini)：GFP和Foxp2-EnhancerA：EGFP。目的2.研究斑马鱼纹状体和MesDA神经元发育过程中轴突的正常分布。我将使用增强子：GFP构建和转基因株系来描述连接性的发展，并将路径发现与Ratio、Sit、Ephs和ePhins的表达模式进行比较。目的3.评价ROBO、SIIT和Ephlephin基因在纹状体和MesDA神经元通路中的作用。我将使用突变鱼线和吗啉的组合来测试这些基因家族中不同成员的功能和效果。临床意义：我们的结果将提高我们对基底节连通性发展的理解，基底节连通性在神经发育和神经退行性疾病中受到影响。摘要：这里描述的工作将是第一次描述基底节的轴突路径，描述连接的发展及其遗传学基础。我的结果将包括基底节发育的描述性元素，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，得到了机构资源的广泛支持和一位国际知名的导师，以帮助我在K奖项期间过渡到一个独立的终身教职跟踪教师职位。

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.