MAPPING NEURAL CIRCUITRY WITH TRANSSYNAPTIC VIRUS

描述（由申请人提供）：神经元形成精确和复杂的神经回路，以产生感知和行为。神经科学的一个主要目标是绘制大脑中神经回路的大集合，并了解它们如何解释正常的大脑功能和神经系统疾病。幼鱼的小尺寸和光学透明度使其成为研究完整脊椎动物生物体神经回路的理想系统。然而，目前用于映射斑马鱼神经回路的技术在速度和规模上都受到限制，阻碍了斑马鱼神经回路研究潜力的充分实现。为了解决这一技术差距，我们建议开发一种基于病毒的工具包，使斑马鱼的神经回路映射快速而系统。在初步研究中，我们确定水泡性口炎病毒（VSV）可用作绘制神经回路的工具。它有效地感染斑马鱼神经元，迅速传播通过突触，并表达荧光报告电路映射。VSV标记也可以与其他成像方法相结合，以探测神经元功能。为了扩展这些研究，我们提出了三个具体目标：目标1。我们将建立基于病毒的工具，在体内绘制全脑规模的神经回路。我们将研究VSV的特征，并将试剂和神经回路映射结果分发给社区。目标2.我们将开发工具来阐明特定神经元类型的连接模式。这些工具将能够（1）将初始感染靶向定义的细胞类型，（2）识别与它们直接连接的神经元。目标3：我们将创建将联合收割机神经电路映射与功能分析相结合的工具。这些工具将有助于弥合神经回路结构和功能之间的差距。总之，这项建议解决了斑马鱼神经回路跟踪工具的迫切需要，并提供了一种新的方法，结合联合收割机解剖和功能分析。我们将利用这些工具来映射视觉功能的电路，既作为一个泰特平台，也作为研究视觉的资源。这些工具可以应用于整个神经系统，并促进更广泛社区的神经回路研究，例如神经发育，生理学，行为，再生以及疾病模型。

DESCRIPTION (provided by applicant): Neurons form precise and complex neural circuits to generate perception and behavior. A major goal of neuroscience is to map the large ensemble of neural circuits in the brain and understand how they account for normal brain functions and neurological disorders. The small size and optical transparency of larval zebrafish make it an ideal system to investigate neural circuits in an intact vertebrate organism. However, current techniques for mapping neural circuitry in zebrafish are limited in speed and scale, preventing the full realization of zebrafish's potential for neural circuit studies. To address this gap in technology, we propose to develop a virus-based toolkit that would enable rapid and systematic neural circuit mapping in zebrafish. In preliminary studies, we established that vesicular stomatitis virus (VSV) can be used as a tool to map neural circuits. It efficiently infects zebrafih neurons, spreads rapidly across synapses, and expresses fluorescent reporters for circuit mapping. VSV labeling could also be combined with other imaging methods to probe neuronal function. To extend these studies, we propose three specific aims: Aim 1. We will establish virus-based tools to map neural circuits on a whole-brain scale, in vivo. We will investigate the characteristics of VSV and distribute reagents and neural circuit mapping results to the community. Aim 2. We will develop tools to elucidate connectivity patterns of specific neuronal types. These tools will be able to (1) target initial infection to defined cell types and (2) identfy neurons that are directly connected to them. Aim 3. We will create tools that combine neural circuit mapping with functional analyses. These tools will help bridge the gap between neural circuit structure and function. In summary, this proposal addresses a critical need for zebrafish neural circuit tracing tools and provides a novel approach to combine anatomical and functional analyses. We will utilize these tools to map the circuitry underlying visual function, both as a tet platform and as a resource for studying vision. These tools could be applied throughout the nervous system and facilitate neural circuit studies in the wider community, e.g. neural development, physiology, behavior, regeneration, as well as disease models.