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复杂且知之甚少的过程。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。虽然一些基因已被鉴定在基底神经节神经元特化或基底神经节疾病中起作用，但这些基因和其他基因如何调节基底神经节连接的发展尚不清楚。此外，在发育过程中基底神经节轴突寻路的实际过程还没有被表征。我的假设是，纹状体和中脑多巴胺能（mesDA）神经元的连接是由robos，狭缝和ephslephrin的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，描述基底神经节连接的发育和遗传控制。为了探索基底神经节连接的发展，本项目有三个具体目标：目标1。开发分子和遗传标记，以可视化纹状体和中脑多巴胺能（mesDA）神经元及其轴突。我用原位基因标记和抗体来标记这些细胞核。为了研究它们的寻路，我已经产生了对基底神经节神经元特异性的新型增强子系，包括dlx（mini）：gfp和foxP 2-enhancerA：egfp。目标2.表征发育过程中斑马鱼纹状体和mesDA神经元的正常轴突寻路。我将使用增强子：绿色荧光蛋白构建体和转基因株系来描述连接性的发展，并将寻路与比率、狭缝、ephs和ephrin的表达模式进行比较。目标3。评估robo、slit和ephlephrin基因在纹状体和mesDA神经元寻路中的作用。我将测试这些基因家族的不同成员的功能和影响，通过使用突变的鱼线和morpholinos组合。临床意义：我们的研究结果将提高我们对基底神经节连通性发展的理解，基底神经节连通性在神经发育和神经退行性疾病中受到影响。总结：这里描述的工作将是第一次描述轴突寻路的基底神经节，表征连接的发展及其遗传基础。我的研究结果将包括基底神经节发育的描述性要素，以及特定基因作用的分析。该项目包括一个结构良好的职业发展计划，广泛支持机构资源和国际知名的导师，以帮助我在K奖期间过渡到一个独立的终身教职。

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.