Role of endocannabinoid signaling in a preference/aversion circuitry

内源性大麻素信号传导在偏好/厌恶电路中的作用

• 批准号: 10365829
• 负责人: Su Guo
• 金额: $ 44.41万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365829
• 关键词: Ablation; Address; Affect; Animal Model; Animals; Anti-Anxiety Agents; Behavior; Behavioral; Brain; Brain region; CNR1 gene; CNR2 gene; Calcium; Cannabinoids; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Corticotropin-Releasing Hormone; Cues; Data; Dependence; Development; Diagnosis; Disease; Drug Addiction; Drug abuse; Elements; Endocannabinoids; Enzymes; Food; Foundations; Frequencies; Functional Imaging; Future; Genes; Genetic; Genetic Models; Glutamates; Head; Human; Hypothalamic structure; Image; Immunologic Receptors; Knock-in; Life; Ligands; Light; Lipids; Mammals; Marijuana; Molecular; Monitor; Motivation; Motor; Motor Activity; Movement; Mus; Neurons; Neuropeptides; Neurotransmitters; Outcome; Pathway Analysis; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phenotype; Policies; Property; Proteins; Public Health; RIPK1 gene; Receptor Inhibition; Regulation; Research; Resolution; Resources; Rewards; Role; Sensory; Shapes; Signal Pathway; Signal Transduction; Slide; Specificity; Stimulus; Stress; Synapses; System; Tail; Testing; Time; Vision; Zebrafish; approach avoidance behavior; avoidance behavior; base; brain cell; cell type; computational platform; drug of abuse; endocannabinoid signaling; exogenous cannabinoid; fitness; genetic regulatory protein; imaging platform; improved; in vivo calcium imaging; interest; knockout animal; motivated behavior; neural circuit; optogenetics; preference; receptor; relating to nervous system; response; sensory stimulus; social; stressor; tool; treatment strategy

PROJECT SUMMARY The ability to seek reward and avoid potential threats is fundamental to the fitness and survival of all animals from early life stages. Our research aims to address circuit-wide mechanisms with cellular and molecular clarity employing larval zebrafish. As a vertebrate genetic model organism, zebrafish shares considerable similarity with mammals. In both mammals and larval zebrafish (See Preliminary data section), the lipid neurotransmitters endocannabinoids (eCB) and the neuropeptide hypothalamic corticotropin releasing factor (Hy CRF) are known to regulate motivated behaviors. However, an understanding of their roles circuit-wide at cellular resolution is currently lacking. Larval zebrafish with a relatively simple and transparent brain of ~100K neurons (compared to ~75 million in the mouse, and ~100 billion in the human brain) is well suited to address this question. New regulatory principles uncovered in simpler systems will lay foundation for studying more complex systems. Free-living with the need to hunt for food and avoid predators, larval zebrafish display readily observable approach and avoidance behaviors in response to environmental stimuli, drugs, or social cues. Here I propose to elucidate the role of eCB and Hy CRF, brain-wide at cellular resolution, in the context of light/dark preference, a fundamental motivated behavior conserved across species. Larval zebrafish avoid dark, which can be enhanced by stressors and alleviated by anxiolytics. Our preliminary data show that ablation of Hy CRF neurons ameliorates, whereas inhibition of the cannabinoid receptor CB1 enhances, dark avoidance. We have genetically disrupted major genes in the eCB signaling pathway, including CB1 (primarily neural) and CB2 (primarily immune) receptors, receptor-interacting proteins (CNRIP1a and CNRIP1b), eCB synthesis enzymes (e.g. DAGLa, DAGLb, ABHD4), and eCB degradation enzymes (MGLL, FAAH). These knockout animals are valuable resources for understanding signaling specificity by uncovering which receptors and ligands and associated regulatory proteins are involved in specific behavioral regulation. Furthermore, we have established brain-wide calcium imaging and computational platforms for examining the activity and plasticity of distributed neural circuits at cellular resolution. In this application, built on these preliminary data and our expertise in studying brain development and function employing zebrafish, we will test the hypothesis that eCB signaling regulates dark avoidance circuitry that involves Hy CRF neurons. We will gain circuit-wide understanding and uncover new cell types/molecules for future studies of circuit assembly and plasticity under stress or drug treatment in a highly accessible brain. Impact and Outcomes: If successful, this project will achieve, for the first time to our knowledge, a cellular resolution circuit-wide understanding of eCB signaling in relation to Hy CRF in a fundamental motivated behavior. Such improved understanding at the whole circuitry level shall lay foundation for informing future marijuana policy and for tackling disease states associated with perturbed CRF or eCB signaling.

项目摘要 寻求奖励和避免潜在威胁的能力是所有动物健康和生存的根本 从生命的早期阶段。我们的研究旨在通过细胞和分子的清晰度来解决电路范围的机制 用的是斑马鱼的幼体斑马鱼作为脊椎动物的遗传模式生物， 哺乳动物。在哺乳动物和斑马鱼幼鱼中（见初步数据部分）， 已知内源性大麻素（eCB）和神经肽下丘脑促肾上腺皮质激素释放因子（Hy CRF 来规范动机行为。然而，了解它们在细胞分辨率中的作用， 目前缺乏。斑马鱼幼鱼具有相对简单和透明的~ 100 K神经元的大脑（比较 在小鼠中约7500万，在人脑中约1000亿）非常适合解决这个问题。新 在简单系统中发现的调节原理将为研究更复杂的系统奠定基础。 自由生活的需要狩猎的食物和避免捕食者，幼斑马鱼显示容易观察 对环境刺激、药物或社会线索的接近和回避行为。在这里我提议 为了阐明eCB和Hy CRF在细胞分辨率方面的全脑作用，在亮/暗偏好的背景下， 一种跨物种的基本动机行为。斑马鱼幼鱼避免黑暗， 由压力源增强并由抗焦虑药缓解。我们的初步数据表明，Hy CRF神经元的消融 改善，而抑制大麻素受体CB 1增强，黑暗回避。从基因上来说 破坏eCB信号通路中的主要基因，包括CB 1（主要是神经）和CB 2（主要是神经）。 免疫）受体、受体相互作用蛋白（CNRIP 1a和CNRIP 1b）、eCB合成酶（例如， DAGLa、DAGLb、ABHD 4）和eCB降解酶（MGLL、FAAH）。这些被击倒的动物很有价值 通过揭示哪些受体和配体以及相关的 调节蛋白参与特定的行为调节。此外，我们还建立了全脑 用于检查分布式神经回路的活动和可塑性的钙成像和计算平台 细胞分辨率。 在这个应用程序中，基于这些初步数据和我们在研究大脑发育和功能方面的专业知识， 使用斑马鱼，我们将测试eCB信号调节黑暗回避电路的假设， 涉及Hy CRF神经元。我们将获得电路范围的理解，并发现新的细胞类型/分子， 未来的研究电路组装和可塑性下的压力或药物治疗在一个高度可及的大脑。 影响和成果：如果成功，这个项目将实现，第一次，我们的知识， 在基本的动机行为中，对与Hy CRF相关的eCB信号传导的解析电路范围的理解。 这种在整个电路水平上的理解的提高将为未来的大麻政策奠定基础 以及用于处理与受干扰的CRF或eCB信号传导相关的疾病状态。