Developing zebrafish models to reveal interactions between stress and addiction

描述(由申请人提供)：通过在分子、细胞和系统水平上产生反应，生命有机体与环境相互作用并适应环境是必不可少的。在某些情况下，这些变化会导致终身甚至几代人的生理和行为变化。了解我们的基因和环境之间的相互作用对于我们理解成瘾至关重要。因此，需要开发分子和遗传工具来剖析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，它由应激反应控制。压力反应是对真实或感知到的威胁或“压力源”做出的生理和/或行为反应，有助于维持或恢复正常的平衡或内稳态。当应激源威胁到动态平衡时，可以利用一套不同的神经元、内分泌和自主神经反应机制来恢复现状。除了对应激源的即时反应外，神经网络或表观基因转录组的长期变化可以记录这一事件的分子记忆，从而改变未来对类似应激源的反应。应激加重障碍的临床意义非常高，当应激源太强(急性)、太频繁(慢性)、或错误地记录到表观基因组和/或在关键的发育时期时，可能会发生。应激反应，尤其是糖皮质激素，会影响成瘾的开始、严重程度、戒断和复发。吸烟和尼古丁成瘾是世界上头号可预防的死亡原因。这项提议将使用遗传模式生物斑马鱼(Danio Rerio)来研究心理刺激剂尼古丁的应激反应和行为敏化的相互作用。具体地说，这项应用将表征暴露于尼古丁后糖皮质激素受体的激活；表征应激源、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感化的变化；以及对应激反应的修饰物进行筛选。这项提议的目标是建立一个数据基础，以加速使用遗传上易驯化的高通量斑马鱼模型来研究压力和成瘾的相互作用。这项赠款的创新要素使用遗传传感器FISH来实现对所有脑组织中糖皮质激素受体激活的时空可视化，以响应药物暴露或其他应激源，并使用基于转座子的插入突变斑马鱼的独特突变库资源来对脊椎动物的应激反应修饰物进行第一次正向遗传筛选。好了！ 与公共卫生相关：压力反应对于适应我们的环境至关重要。不幸的是，压力可以影响成瘾过程的许多方面，包括开始、严重、戒断和复发。对像斑马鱼这样的遗传易驯化的高通量模式脊椎动物的适应，可能会导致发现新的遗传途径，这些途径有助于对成瘾和压力的脆弱性或弹性。

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.