Imaging tools to assess circuit connectivity and function in zebrafish

项目摘要 神经元 发展 神经元 组成 协调 理解 人口 建议， 运动前 传感器 复溶 他们的 的 的 标记 神经元 interneurons 是 神经回路产生于突触的活动依赖性形成和消除， 活动依赖性突触连接在外周感觉和运动神经中已被广泛研究 目标是兴奋或兴奋。然而，在中枢神经系统中，神经回路 兴奋和抑制共同目标的神经元，因此活动依赖性布线必须 在发展过程中，以达到适当的平衡的兴奋和抑制。更好地 在这个过程中，它将有助于同时监测兴奋性和抑制性的活动， 以及它们在发育过程中与共同目标的突触连接。在这个探索性的R21中， 我们将开发一种遗传策略来监测兴奋性和抑制性神经元的活动依赖性变化， 发育中的斑马鱼脊髓中的突触连接。我们将使用一种基因编码的 在黑腹果蝇中开发，使用基于小突触蛋白的GFP、YFP或CFP 同时用不同的颜色标记连接， 活性水平（Syb-XRASP）。在目标1中，我们将验证Syb-XRASP的CFP、YFP和GFP变体， 发育中的斑马鱼的神经肌肉接头。在Aim 2中，我们将使用Syb-XRASP来监控组件 兴奋性Chx 10标记的V2 a群和抑制性En 1- V1人群。总之，这些目标将使我们能够评估从运动神经元连接的形成。 从单个V2 a和V1中间神经元到所有运动神经元，从所有V2 a和V1中间神经元到肌肉， 到单个运动神经元。这些实验将提供一个前所未有的视角， 在由兴奋性和抑制性神经元组成的回路发育过程中形成。 t t F

Project Summary Neuronal development. neurons comprised coordinated understand populations proposal, premotor sensor reconstitution their the of labeled neurons interneurons are circuits arise from the activity-dependent formation and elimination of synapses during Activity-dependent synaptic wiring has been studied extensively in peripheral sensory and motor where the goal is o be excited or o excite. However, in the central nervous system, neural circuits are of neurons that both excite and inhibit common targets and so activity-dependent wiring must be during development to achieve an appropriate balance of excitation and inhibition. To better this process, it would help to simultaneously monitor the activity of excitatory and inhibitory and their synaptic connections to common targets during development. In this exploratory R21 we will develop a genetic strategy to monitor activity-dependent changes in excitatory and inhibitory synaptic connectivity in the developing zebrafish spinal cord. We will use a genetically-encoded developed in Drosophila melanogaster that uses a synaptobrevin-based GFP, YFP or CFP across synaptic partners to simultaneously label connections in different colors and monitor activity levels (Syb-XRASP) . In Aim 1 we will validate the CFP, YFP and GFP variants of Syb-XRASP at neuromuscular junction of developing zebrafish. In Aim 2, we will use Syb-XRASP to monitor the assembly connections of premotor neurons within excitatory Chx10-labeled V2a population and inhibitory En1- V1 population. Together, these aims will allow us to assess the formation of connections rom motor to muscles, from single V2a and V1 i nterneurons to all motor neurons, and from all V2a and V1 to single motor neurons. These experiments will provide an unprecedent view of how connections formed during the development of circuits comprised of excitatory and inhibitory neurons. t t f