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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要确定大脑中神经元如何相互连接是迈向这一目标的重要且必要的一步了解大脑活动如何产生行为，以及它如何受到疾病的干扰。目前可用的方法存在局限性，使得可视化这些大脑接线图具有挑战性。此外，迫切需要一种方法，不仅能够揭示大脑连接性，而且能够揭示大脑连接性。我们最近开发了一种对连接在电路中的神经元的功能特性进行基因改造的方法。名为 TRACT 的遗传系统并用果蝇证明它具有这两种特征。许多复杂的大脑功能无法在果蝇中进行检查，了解它们需要使用近年来，斑马鱼已成为研究有用的脊椎动物模型。由于其相对简单但保守的脊椎动物大脑，复杂的大脑过程，光学透明胚胎和幼虫发育阶段、大规模行为分析的适应性、仅仅 5 天的发育后就出现了复杂的行为，并且越来越多的遗传工具可以进行观察然而，斑马鱼在神经科学中的用途。研究因缺乏识别和扰乱突触连接神经元的方法而受到限制。初步研究，我们开发了一个TRACT系统，可用于识别顺行单突触在这里，我们建议进一步开发 TRACT 作为一种工具。通过开发额外的解剖追踪模式并建立斑马鱼的跨神经元追踪使用 TRACT 对突触连接的神经元进行遗传操作和功能表征。功能将增加斑马鱼作为研究脊椎动物神经回路的模型系统的有用性功能，既揭示特定行为背后的神经回路的一般原理，又建模复杂的脑部疾病，如自闭症、阿尔茨海默病和精神分裂症。