Dynorphinergic projections from the insular cortex to the central amygdala: role in stress-enhanced alcohol drinking in dependence

项目摘要 酒精使用障碍（AUD）是一种流行的慢性复发性疾病，其中压力起着重要作用。 了解压力和饮酒之间相互作用的神经生物学基础对于 打击澳元的发展和维护。中央杏仁核（CeA）在 调节酒精相关的行为，特别是在酒精依赖的情况下，并对压力作出反应。 类似地，岛叶皮层（IC）已被证明可以调节滥用药物的渴望和寻求行为， 包括酒精，也与情绪处理有关。强啡肽/κ阿片受体（Dyn/KOR） 信号已经被证明编码负面影响，就像压力和酒精引起的那样 撤回，和Dyn/KOR在IC和CeA中均稳健表达。此外，Dyn/KOR的中断 已经显示CeA中的信号传导阻断酒精依赖小鼠中的应激增强的酒精饮用。我们 有初步证据表明，IC向CeA发送了一个大的Dyn表达（Dyn+）投射， 介导这种效应。据我们所知，迄今为止还没有研究检查过这种ICDyn+ ICDceA回路，以及它是如何 规范过度饮酒。因此，拟议的研究将描述这种电路，研究如何 长期暴露在压力和酒精中会影响回路的活动，以及操纵这个回路会如何影响 酒精依赖小鼠的应激增强饮酒。我们的首要假设是， ICDyn+ ICD 依赖为了验证这一假设，我们将使用Stress-CIE饮酒模型，这是一种小鼠模型， 慢性间歇性乙醇（CIE）暴露与慢性强迫游泳应激（FSS）。这个模型产生酒精 依赖和增加酒精摄入与压力。对于目标1，我们将表征 使用病毒追踪和免疫荧光技术在压力-CIE饮酒模型中的ICDyn+ ICDceA回路。为 目的2：采用纤维光度法测定笼内饮酒时ICDyn+ β CeA环路的活性 在Stress-CIE饮酒模型中。对于目标3，将使用Stress-CIE饮酒模型结合设计师 受体专门激活的设计师药物（DREADDs），以操纵ICDyn+ ICDyn CeA电路在家里 笼子里喝酒总的来说，这些研究将阐明研究不足的ICDyn+ ICDceA电路的作用。 以及它如何在酒精依赖和酒精压力相互作用的模型中调节饮酒。

PROJECT SUMMARY Alcohol use disorder (AUD) is a prevalent, chronically relapsing disorder in which stress plays a major role. Understanding the neurobiological underpinnings of interactions between stress and alcohol drinking is vital to combating the development and maintenance of AUD. The central amygdala (CeA) plays an important role in regulating alcohol-related behaviors, especially in the context of alcohol dependence, and is responsive to stress. Similarly, the insular cortex (IC) has been shown to regulate craving and seeking behaviors with drugs of abuse, including alcohol, and is also implicated in emotional processing. Dynorphin/kappa opioid receptor (Dyn/KOR) signaling has been shown to encode negative affect, like that which is caused during stress and alcohol withdrawal, and Dyn/KOR are robustly expressed in both the IC and CeA. Furthermore, disruption of Dyn/KOR signaling in the CeA has been shown to block stress-enhanced alcohol drinking in alcohol dependent mice. We have preliminary evidence that the IC sends a large Dyn-expressing (Dyn+) projection to the CeA that may mediate this effect. To our knowledge, no studies to date have examined this ICDyn+CeA circuit and how it regulates excessive alcohol drinking. Thus, the proposed studies will characterize this circuit, examine how chronic exposure to stress and alcohol affects the circuit's activity, and how manipulation of this circuit affects stress-enhanced alcohol drinking in alcohol dependent mice. Our overarching hypothesis is that the ICDyn+CeA circuit is a critical mediator of stress-enhanced alcohol drinking in the context of dependence. To test this hypothesis, we will use the Stress-CIE Drinking model, a mouse model that combines chronic intermittent ethanol (CIE) exposure with chronic forced swim stress (FSS). This model produces alcohol dependence and enhanced alcohol intake with stress. For Aim 1, we will characterize the activity of the ICDyn+CeA circuit in the Stress-CIE Drinking model using viral tracing and immunofluorescence techniques. For Aim 2, fiber photometry will be used to measure ICDyn+CeA circuit activity during home cage alcohol drinking in the Stress-CIE Drinking model. For Aim 3, will use the Stress-CIE Drinking model combined with designer receptors exclusively activated by designer drugs (DREADDs) to manipulate the ICDyn+CeA circuit during home cage alcohol drinking. Collectively, these studies will shed light on the role of the understudied ICDyn+CeA circuit and how it regulates alcohol drinking in a model of alcohol dependence and alcohol-stress interactions.