Mapping Neural Connectivity in Zebrafish Larvae Using a Photoconvertible Protein

使用光转换蛋白绘制斑马鱼幼虫的神经连接图

• 批准号: 10437658
• 负责人: Adam Christopher Roberts
• 金额: $ 11.11万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10437658
• 关键词: Affect; Alzheimer' s Disease; Animal Model; Animals; Base of the Brain; Behavior; Behavioral; Biological; Biological Models; Brain; Brain Diseases; Brain Mapping; Caenorhabditis elegans; Data; Development; Disease; Drug Addiction; Drug Screening; Etiology; Extracellular Signal Regulated Kinases; Fertility; Fishes; Functional disorder; Goals; Human; Invertebrates; Investigation; Larva; Learning; MAP Kinase Gene; Maps; Measures; Memory; Mental disorders; Methods; Modeling; Modernization; Modification; Molecular; Mus; Neurodevelopmental Disorder; Neurological Models; Neuronal Plasticity; Neurons; Neurosciences; Organism; Pathology; Physiological; Population; Post-Traumatic Stress Disorders; Procedures; Property; Proteins; Reporting; Research Personnel; Schizophrenia; Signal Transduction; System; Technology; Time; Transgenic Organisms; United States National Institutes of Health; Vertebrates; Zebrafish; autism spectrum disorder; calcium indicator; connectome; effective therapy; experimental study; high throughput screening; human model; in vivo; interest; nervous system disorder; neural circuit; neuropsychiatric disorder; novel therapeutics; promoter; relating to nervous system; response; sensory system; therapy development; tool; usability

PROJECT SUMMARY Creation of whole-brain functional connectomes will facilitate a holistic understanding of memory formation, which is a major goal of modern neuroscience. Information gained from understanding the functional connections between neurons and the underlying plasticity between connections can be used to understand and treat neurological and mental disorders. In vivo whole-brain mapping with current technologies requires advantageous biological attributes in combination with a substantial molecular toolbox. Two invertebrate model organisms have these properties: D. melanogaster and C. elegans. However, zebrafish (Danio rerio) is the only vertebrate species with this capacity. The ability to form a functional connectome with larval zebrafish affords distinct advantages to understanding memory formation or pinpointing aberrant neural circuits in animal models of human disorders. Further, rapid development, small size, and high fecundity of zebrafish makes them an ideal organism for high- throughput screening, a useful mechanism to discover novel therapeutics for these disorders. The project will attempt to create a new tool to rapidly form in vivo whole-brain connectomes in freely moving fish with the long-term goal of understanding brain function and dysfunction at a circuit-level. Towards this goal, we will create transgenic fish that express a photoconvertible protein under the control of activity- dependent promoters. We will then determine the experimental procedures for each line of transgenic fish that most efficiently and accurately reflect the neural activity under investigation and minimize signals related to nonspecific neural activity. After we have defined the experimental potential and constraints of our transgenic lines of fish, we will seek to determine if the recorded neural activity is physiologically valid. To do so, we will form functional maps of neural activity in response to stimulation of sensory systems with previously defined neural connections. These experiments should provide the pilot data necessary to begin investigating changes in functional connectomes due to memory formation in zebrafish models of neurodevelopmental disorders. Data from the proposed studies are expected to facilitate the development of effective treatments for brain diseases and disorders, including Alzheimer's disease, PTSD, schizophrenia and autism.

项目概要 创建全脑功能连接组将有助于对记忆的整体理解 形成，这是现代神经科学的一个主要目标。从理解函数中获得的信息 神经元之间的连接以及连接之间潜在的可塑性可以用来理解 并治疗神经和精神疾病。 使用当前技术进行体内全脑图谱需要有利的生物属性 与大量的分子工具箱相结合。两种无脊椎动物模式生物具有以下特性：D. 黑腹果蝇和线虫。然而，斑马鱼（Danio rerio）是唯一具有此功能的脊椎动物物种 容量。与斑马鱼幼虫形成功能性连接组的能力为 了解记忆形成或查明人类疾病动物模型中的异常神经回路。 此外，斑马鱼发育快、体型小、繁殖力强，使其成为高产的理想生物。 通量筛选是发现这些疾病的新疗法的有用机制。 该项目将尝试创建一种新工具，能够在体内自由地快速形成全脑连接组。 移动鱼的长期目标是在电路层面了解大脑功能和功能障碍。向 为了实现这个目标，我们将创造出在活性控制下表达光转换蛋白的转基因鱼—— 依赖性启动子。然后我们将确定每个转基因鱼品系的实验程序 最有效、最准确地反映正在研究的神经活动，并最大限度地减少与相关的信号 非特异性神经活动。在我们确定了转基因的实验潜力和限制之后 鱼线，我们将设法确定记录的神经活动在生理上是否有效。为此，我们将 形成神经活动的功能图，以响应先前定义的感觉系统的刺激 神经连接。这些实验应该提供开始调查变化所需的试点数据 由于神经发育障碍的斑马鱼模型中的记忆形成而导致功能性连接组的变化。 拟议研究的数据预计将有助于开发有效的治疗方法 脑部疾病和紊乱，包括阿尔茨海默病、创伤后应激障碍、精神分裂症和自闭症。