MAPPING NEURAL CIRCUITRY WITH TRANSSYNAPTIC VIRUS

用跨突触病毒绘制神经回路

• 批准号: 8889264
• 负责人: YUCHIN Albert Pan
• 金额: $ 37.24万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8889264
• 关键词: Accounting; Address; Affect; Anatomy; Animal Behavior; Architecture; Autistic Disorder; Behavior; Blindness; Brain; Brain Mapping; Brain imaging; Brain region; Calcium; Cells; Characteristics; Collection; Communities; Complex; Data; Detection; Development; Disease model; Embryo; Environment; Functional Imaging; Genetic; Goals; Health; Human; Image; Infection; Injury; Kinetics; Knowledge; Label; Larva; Lead; Maps; Mediating; Membrane; Methods; Movement; Natural regeneration; Nervous system structure; Neurons; Neurosciences; Optics; Organ; Organism; Parkinson Disease; Pattern; Perception; Physiological; Physiology; Population; Process; Psyche structure; Reagent; Reporter; Resolution; Resources; Retinal Ganglion Cells; Speed; Spinal cord injury; Structure; Synapses; System; Techniques; Technology; Testing; Tetanus Helper Peptide; Tissues; Transgenic Organisms; Vesicular stomatitis Indiana virus; Viral; Virus; Virus Diseases; Vision; Visual; Visual Motion; Zebrafish; base; behavioral study; cell type; functional loss; human disease; imaging modality; in vivo; insight; nervous system disorder; neural circuit; neurodevelopment; novel; novel strategies; prevent; receptor; relating to nervous system; response; tool; transmission process; vector

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.

描述（由申请人提供）：神经元形成精确而复杂的神经回路，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量合奏，并了解它们如何解释正常的大脑功能和神经系统疾病。幼虫斑马鱼的尺寸较小和光学透明度使其成为研究完整脊椎动物中神经回路的理想系统。但是，斑马鱼中映射神经回路的当前技术的速度和尺度有限，从而完全实现了斑马鱼的神经回路研究潜力。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，该工具包可以在斑马鱼中快速而系统的神经电路映射。在初步研究中，我们确定囊泡口腔炎病毒（VSV）可以用作绘制神经回路的工具。它有效地感染了斑马神经元，迅速传播在突触中，并表达用于电路映射的荧光记者。 VSV标记也可以与其他成像方法结合使用，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。我们将在体内建立基于病毒的工具，以全脑尺度绘制神经回路。我们将研究VSV的特征，并将试剂和神经电路映射结果分配给社区。目标2。我们将开发工具来阐明特定神经元类型的连通性模式。这些工具将能够（1）将初始感染靶向定义的细胞类型，以及（2）直接连接到它们的识别神经元。 AIM 3。我们将创建将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电路结构与功能之间的差距。总而言之，该提案解决了斑马鱼神经电路追踪工具的关键需求，并提供了一种结合解剖学和功能分析的新方法。我们将利用这些工具将视觉功能的基础电路绘制为TET平台和研究视觉的资源。这些工具可以在整个神经系统中应用，并促进更广泛的社区的神经回路研究，例如神经发育，生理，行为，再生以及疾病模型。