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

基本信息

批准号：
10608111
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
项目类别：
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/10608111
关键词：
Ablation Address Affect Animals Anti-Anxiety Agents Behavior Behavioral Brain Brain region CNR1 gene CNR2 gene Calcium Cannabinoids Cannabis policy Classification Clustered Regularly Interspaced Short Palindromic Repeats Complex Corticotropin-Releasing Hormone Cues Darkness 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 Immune Knock-in Life Ligands Light Lipids Mammals Molecular Monitor Motivation Motor Motor Activity Movement Mus Neurons Neuropeptides Neurotransmitters Outcome Pathway Analysis Pharmaceutical Preparations 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 System Tail Testing Time Vision Zebrafish approach behavior avoidance behavior 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 model organism motivated behavior neural neural circuit optogenetics pharmacologic postsynaptic preference receptor response sensory stimulus social stressor tool treatment strategy

项目摘要

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.

项目概要寻求奖励和避免潜在威胁的能力是所有动物的健康和生存的基础从生命的早期阶段。我们的研究旨在以细胞和分子的清晰度解决全电路机制使用斑马鱼幼体。作为脊椎动物遗传模式生物，斑马鱼具有相当大的相似性与哺乳动物。在哺乳动物和斑马鱼幼虫中（参见初步数据部分），脂质神经递质已知内源性大麻素 (eCB) 和神经肽下丘脑促肾上腺皮质激素释放因子 (Hy CRF) 来调节动机行为。然而，在细胞分辨率下了解它们在整个电路中的作用目前缺乏。斑马鱼幼虫的大脑相对简单且透明，约有 10 万个神经元（与小鼠大脑中约为 7500 亿，人脑中约为 1000 亿）非常适合解决这个问题。新的在更简单的系统中发现的监管原则将为研究更复杂的系统奠定基础。斑马鱼幼体自由生活，需要寻找食物并躲避捕食者，其表现很容易观察到对环境刺激、药物或社交线索的接近和回避行为。在这里我建议阐明 eCB 和 Hy CRF 在全脑细胞分辨率下在光/暗偏好背景下的作用，跨物种保守的基本动机行为。斑马鱼幼体避免黑暗，这可能是应激源增强，抗焦虑药缓解。我们的初步数据表明 Hy CRF 神经元的消融改善黑暗回避，而抑制大麻素受体 CB1 则增强回避黑暗的能力。我们有遗传基因破坏了 eCB 信号通路中的主要基因，包括 CB1（主要是神经）和 CB2（主要是神经）免疫）受体、受体相互作用蛋白（CNRIP1a 和 CNRIP1b）、eCB 合成酶（例如 DAGLa、DAGLb、ABHD4) 和 eCB 降解酶（MGLL、FAAH）。这些淘汰动物很有价值通过揭示哪些受体和配体以及相关的来了解信号特异性的资源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在细胞分辨率下。在此应用程序中，基于这些初步数据以及我们在研究大脑发育和功能方面的专业知识使用斑马鱼，我们将测试以下假设：eCB 信号调节暗回避电路涉及 Hy CRF 神经元。我们将获得全电路的理解并发现新的细胞类型/分子未来的研究是在高度可及的大脑中进行压力或药物治疗下的电路组装和可塑性。影响和成果：据我们所知，如果成功，该项目将首次实现细胞在基本动机行为中解决与 Hy CRF 相关的 eCB 信号传导的全电路理解。这种对整个电路层面的更好的理解将为未来的大麻政策奠定基础用于解决与 CRF 或 eCB 信号传导紊乱相关的疾病状态。