Developing zebrafish models to reveal interactions between stress and addiction

描述（由申请人提供）：生物通过在分子、细胞和系统水平上产生反应来与环境相互作用和适应环境是必不可少的。在某些情况下，这些变化会导致一生甚至几代人的生理和行为变化。了解基因和环境之间的相互作用对我们理解成瘾至关重要。因此，需要开发分子和遗传工具来解剖和发现这些相互作用。至关重要的是，基因和行为之间的桥梁是由应激反应控制的。应激反应是对真实或感知到的威胁或“压力源”的生理和/或行为反应，有助于维持或恢复正常的平衡或体内平衡。当压力源威胁到体内平衡时，可以利用多种神经元、内分泌和自主反应机制来恢复现状。除了对应激源的即时反应外，神经网络或表观遗传转录组的长期变化可以记录该事件的分子记忆，从而改变未来对类似应激源的反应。应激加重性疾病的临床意义非常高，当应激源太强（急性）、发生太频繁（慢性）、或不正确地记录到表观基因组和/或在关键发育时期时，可能发生应激加重性疾病。应激反应，特别是糖皮质激素，影响成瘾的发作、严重程度、戒断和复发。吸烟以及由此产生的尼古丁成瘾，是世界上可预防的头号死因。本提案将使用遗传模式生物斑马鱼（Danio rerio）来研究应激反应和对精神兴奋剂尼古丁的行为敏感的相互作用。具体地说，这种应用将表征暴露于尼古丁后糖皮质激素受体的激活；描述由应激源、外源性皮质类固醇或应激反应拮抗剂引起的尼古丁行为敏化的变化；并对压力反应的调节器进行筛选。本提案的目标是建立一个数据基础，以加速使用遗传易学，高通量的斑马鱼模型来研究压力和成瘾的相互作用。这项资助的创新元素使用遗传传感器鱼来实现糖皮质激素受体在所有脑组织中对药物暴露或其他压力源的响应的时空可视化，以及基于转座子插入突变的斑马鱼的独特突变文库资源，以进行脊椎动物应激反应修饰因子的首次前向遗传筛选。！

DESCRIPTION (provided by applicant): It is essential for a living organism to interact with and adjust to the environment by generating responses at molecular, cellular, and system levels. In some cases, these changes result in life-long or even multi- generational changes to physiology and behavior. Understanding the interactions between our genes and the environment is vital to our understanding of addiction. Therefore, molecular and genetic tools to dissect and discover these interactions need to be developed. Of critical importance is the bridge between genetics and behavior that is gated by the stress response. A stress response is a physiological and/or behavioral response to a real or perceived threat or "stressor" that helps maintain or restore normal balance or homeostasis. When a stressor threatens homeostasis, a diverse suite of neuronal, endocrine, and autonomic response mechanisms can be utilized to regain the status quo. In addition to the immediate changes in response to a stressor, long-term changes in the neural network or epigenetic transcriptome can record the molecular memory of this event and thereby alter future responses to similar stressors. The clinical significance of stress-aggravated disorders is extremely high, and can occur when a stressor is too strong (acute), occurs too often (chronic), or is recorded to the epigenome incorrectly and/or during critical developmental periods. The stress response, particularly glucocorticoids, affects the onset, severity, withdrawal, and relapse of addiction. Smoking and thereby nicotine addiction, is the number one preventable cause of death in the world. This proposal will use a genetic model organism, zebrafish (Danio rerio), to study the interaction of the stress response and behavioral sensitization to a psychostimulant- nicotine. Specifically this application will characterize glucocorticoid receptor activation following exposure to nicotine; characterize changes in behavioral sensitization to nicotine caused by stressors, exogenous corticosteroids, or antagonists to the stress response; and perform a screen for modifiers of the stress response. The goal of this proposal is to build a foundation of data to accelerate the use of the genetically tractable, high-throughput zebrafish model to study the interaction of stress and addiction. Innovative elements of this grant use genetic sensor fish to permit spatio-temporal visualization of glucocorticoid receptor activation in all brain tissues in response to drug exposure or other stressors, and a unique mutant library resource of transposon-based insertional mutant zebrafish to do the first forward genetic screen for modifiers of the stress response in a vertebrate. ! PUBLIC HEALTH RELEVANCE: The stress response is vital for adaptation to our environment. Unfortunately, stress can influence many aspects of the addiction process including onset, severity, withdrawal, and relapse. The adaptation of a genetically tractable, high throughput model vertebrate organism, like zebrafish, could lead to the discovery of new genetic pathways that contribute to vulnerability or resilience to addiction and stress.