Development and Validation of a Genetically Encoded Method to Trace and Manipulate Neuronal Circuits in Zebrafish

追踪和操纵斑马鱼神经元回路的基因编码方法的开发和验证

• 批准号: 10505822
• 负责人: CARLOS LOIS
• 金额: $ 244.05万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-19 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10505822
• 关键词: Ablation; Address; Adopted; Adult; Affect; Alzheimer' s Disease; Anatomy; Animal Model; Animals; Behavior; Behavioral; Behavioral Assay; Biological Models; Brain; Brain Diseases; Cerebellum; Collection; Communities; Complex; Development; Disease; Drosophila genus; Due Process; Embryo; Ensure; Gene Expression; Genes; Genetic; Injections; Interneurons; Link; Methods; Modality; Modeling; Modification; Names; Neurons; Neurosciences; Neurosciences Research; Optics; Population; Process; Property; Proteins; Reporter Genes; Reproducibility; Schizophrenia; Sleep; Synapses; System; Technology; Transgenic Organisms; Validation; Variant; Vertebrates; Virus; Visual; Zebrafish; anatomical tracing; autism spectrum disorder; calcium indicator; experimental study; genetic manipulation; improved; interest; motor learning; neuronal circuitry; neuropsychiatric disorder; new technology; postsynaptic neurons; presynaptic; public health relevance; reconstitution; tool

PROJECT SUMMARY Identifying how neurons are connected to each other in the brain is an important and necessary step towards understanding how brain activity gives rise to behavior, and how it is perturbed by disease. Unfortunately, currently available methods have limitations that make it challenging to visualize these brain wiring diagrams. In addition, there is an urgent need for a method that will make it possible not only to unveil brain connectivity, but also to genetically modify the functional properties of neurons connected in a circuit. We recently developed a genetic system named TRACT and showed using Drosophila that it possesses both of these features. However, many complex brain functions cannot be examined in Drosophila, and understanding them will require the use of vertebrate animals. In recent years the zebrafish has emerged as a useful vertebrate animal model to study complex brain processes due to its relatively simple yet conserved vertebrate brain, optical transparency during embryonic and larval stages of development, amenability to large-scale behavioral assays, the emergence of complex behaviors after only 5 days of development, and a growing suite of genetic tools that allow observation and manipulation of neuronal circuits in behaving animals. However, the usefulness of zebrafish for neuroscience research is constrained by a lack of methods to identify and perturb synaptically connected neurons. In preliminary studies, we developed a TRACT system that can be used to identify anterograde monosynaptic connections between neurons in the zebrafish brain. Here we propose to further develop TRACT as a tool for transneuronal tracing in zebrafish by developing additional anatomical tracing modalities, and by establishing the use of TRACT to genetically manipulate and functionally characterize synaptically connected neurons. These capabilities will increase the usefulness of zebrafish as a model system to study vertebrate neuronal circuit function, both to reveal general principles of neuronal circuits that underlie specific behaviors, and to model complex brain disorders such as autism, Alzheimer’s disease and schizophrenia.

项目摘要 确定神经元如何在大脑中相互联系是迈向的重要和必要的步骤 了解大脑活动如何引起行为以及疾病如何扰动它。很遗憾， 当前可用的方法有局限性，使得可视化这些大脑接线图的挑战。在 此外，迫切需要一种方法，该方法不仅可以揭示大脑连接性，而且还可以 我们最近开发了一个 遗传系统命名为“小道”，并使用果蝇具有这两种特征。然而， 许多复杂的大脑功能无法在果蝇中检查，理解它们将需要使用 脊椎动物。近年来，斑马鱼已成为一种有用的脊椎动物模型来研究 复杂的大脑过程由于其相对简单但保守的脊椎动物大脑，光学透明度 发育的胚胎和幼虫阶段，对大规模行为测定的适用性，出现 仅在开发5天之后，复杂的行为以及越来越多的遗传工具允许观察 以及在行为动物中操纵神经元电路。但是，斑马鱼对神经科学的有用性 研究受到缺乏识别和扰动突触连接神经元的方法的限制。 初步研究，我们开发了一种可用于识别稳定单突触的区域 斑马鱼大脑中神经元之间的连接。在这里，我们建议进一步开发作为一种工具 通过开发额外的解剖学追踪方式，并建立透脑性的追踪 使用道的使用来一般操纵并在功能上表征突触连接的神经元。 功能将增加斑马鱼作为研究脊椎动物神经元电路的模型系统的实用性 功能，既揭示了基于特定行为的神经元电路的一般原理，又要建模 复杂的脑疾病，例如自闭症，阿尔茨海默氏病和精神分裂症。