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

项目总结 酒精使用障碍(AUD)是一种普遍存在的慢性复发性障碍，压力在其中起主要作用。 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 调节与酒精有关的行为，特别是在酒精依赖的情况下，并对压力做出反应。 同样，岛叶皮质(IC)已被证明通过滥用药物来调节渴望和寻找行为， 包括酒精，而且还牵涉到情绪处理。强啡肽/kappa阿片受体(Dyn/KOR) 信号已被证明可以编码负面影响，就像在压力和酒精中产生的那样 撤退和Dyn/KOR在IC和CEA中都有强有力的表达。此外，Dyn/KOR的中断 CEA中的信号已被证明可以阻止酒精依赖小鼠应激增强的饮酒。我们 有初步证据表明IC向CEA发送了一个大的dyn表达(dyn+)投影，这可能 调停这一效应。据我们所知，到目前为止还没有研究检查ICDyn+CEA电路以及它是如何 规定过度饮酒。因此，拟议的研究将描述这一电路，检查如何 长期暴露在压力和酒精中会影响该回路的活动，以及对该回路的操纵如何影响 酒精依赖小鼠的压力增强型饮酒。我们最重要的假设是 ICDyn+CEA回路是压力增强型饮酒的关键调节因子 依赖。为了验证这一假设，我们将使用压力-CIE饮酒模型，这是一种结合了 慢性间歇性酒精(CIE)暴露与慢性强迫游泳应激(FSS)。这种型号可以生产酒精。 依赖和增加压力下的酒精摄入量。对于目标1，我们将描述 应用病毒示踪和免疫荧光技术在应激-CIE饮水模型中建立ICDyn+癌胚抗原回路。为 目的2、采用光纤光度法检测家庭笼养饮酒过程中ICDyn+癌胚抗原环路活性 在压力-CIE饮酒模型中。对于目标3，将使用压力-CIE饮酒模式与设计师相结合 专门由特制药物(DREADD)激活的受体在家中操纵ICDyn+CEA电路 笼子里的酒精饮料。总的来说，这些研究将阐明未被研究的ICDyn+CEA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酒精饮酒。

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.