Insula to Limbic circuits differentially mediate BHID expression in alcohol-preferring rodent models

项目摘要/摘要 酗酒是酒精使用障碍(AUD)危害的一个主要因素，高强度的酗酒 摄入是特别危险的。然而，人们对驱动前载的机制(S)知之甚少 (强劲的初始摄入量)和持续的高饮酒率。第一部分人类结果表明，家族性 AUD的病史/密度与达到暴饮暴食水平的时间较短有关，提示过度饮酒的可遗传风险 饮酒过量的原因很简单，但仍有严重的未得到满足的需要需要了解过度饮酒的驱动机制(S)。至 发现关键的大脑回路，驱动过量摄入并具有遗传风险，我们使用高风险选择的大鼠品系 饮酒，P和HAD大鼠，这可能是遗传性AUD风险的模型，以及适度饮酒的Wistar大鼠 作为对照。重要的是，P鼠表现出更大的行为反应，包括刺激的突出性和活力，而 让老鼠有更高的焦虑感。因此，这两条线可能是澳元奖励饮酒与救济饮酒的典范。 过度饮酒很可能是由酒精强烈而显著的动机特性驱动的，这意味着 大脑的显著网络(SN)，它识别并快速响应重要事件。我们专注于前部 脑岛(ANS)，SN的关键调节者，以及动机和情绪驱动行为的许多方面。另外， 抑制AINS减少啮齿动物饮酒，人类研究表明AINS与暗示和否定有关 受情感驱使的酒精驱动力。然而，人们对ANS的具体机制知之甚少(S) 促进前置负荷和持续高饮酒。我们专门研究了AINS对原子核的投射 伏隔核(NAcb)和基底外侧杏仁核(BLA)，因为NAcb被广泛地与活力和 奖赏导向的行为，与焦虑表达有关的血乳酸。因此，我们假设P鼠饮酒将 在饮酒时涉及更多的INS-NAcb神经元放电，在抑制时更大程度地减少消耗 AINS-NAcb(VS HAD和Wistar大鼠)。相比之下，HADS将有更高的饮酒和BLA编码 当抑制ANS-BLA时，摄入量减少更多。目标1寻找大鼠在解剖尺寸上的品系差异 AINS-NAcb和AINS-BLA预测。AIM 2使用最先进的活体神经元记录来发现放电 AINS-NAcb和AINS-BLA神经元在前负荷期间的模式(通过光标记识别，基于光 识别特定细胞的刺激)。我们预测，更大的AIN发射将与更高的前置负荷相关， 尤其是HAD大鼠的AINS-BLA和P大鼠的AINS-NACORE。在饮酒之前，我们还检查了在饮酒过程中 焦虑、镇静和蔗糖暴露，以确定这些状态的编码是否可以预测 摄入量过高。AIM 3使用投影特异性抑制方法特异性抑制Ains-NAcore 或INS-BLA，并检验过量饮酒更受INS-NAcore抑制的影响的假设 在P大鼠和在HADS中抑制INS-BLA。总之，拟议的研究具有很强的互动性和协同性。 对于其他组件，如果成功，我们工作的积极影响将是发现关键的AN 驱动前负荷和持续高酒精饮酒的机制，这将改善AUD的治疗。

Project Summary/Abstract Binge drinking is a major contributor to the harms of alcohol use disorder (AUD), with high-intensity binge intake being particularly hazardous. However, little is known about the mechanism(s) that drive front-loading (strong initial intake rate) and sustained high drinking. Component 1 human results show that familial history/density of AUD relates to shorter time to reach binge levels, suggestive a heritable risk for excessive intake, but there is critical unmet need to understand mechanism(s) that drive excessive drinking. To discover critical brain circuits that drive excessive intake with heritable risk, we use rat lines selected for high alcohol consumption, P and HAD rats, which may model heritable AUD risk, and moderate-drinking Wistar rats as controls. Importantly, P rats show greater behavioral responses involving stimulus salience and vigor, while HAD rats have higher anxiety. Thus, these two lines may model reward versus relief drinking in AUD. Excessive drinking is likely driven by the strong, salient motivational properties of alcohol, which implicates the brain’s salience network (SN) which identifies and rapidly responds to important events. We focus on anterior insula (aINS), a critical regulator of SN and many aspects of motivation- and emotion-driven behavior. Also, aINS inhibition reduces rodent alcohol drinking, and human studies implicate aINS in both cue- and negative affect-driven drive for alcohol. However, little is known about specific mechanism(s) whereby aINS promotes front-loading and sustained high drinking. We specifically examine aINS projections to Nucleus Accumbens core (NAcb) and Basolateral Amygdala (BLA), since NAcb has widely been linked to vigor and reward-directed action, with BLA implicated in anxiety expression. Thus, we hypothesize that P rat drinking will involve greater aINS-NAcb neuronal firing during drinking and greater reduction of consumption when inhibiting aINS-NAcb (vs HAD and Wistar rats). In contrast, HADs will have higher aINS-BLA encoding of drinking and greater reduction of intake when inhibiting aINS-BLA. Aim 1 looks for rat line differences in the anatomical size of aINS-NAcb and aINS-BLA projections. Aim 2 uses cutting edge in vivo neuronal recording to discover firing patterns during front-loading in aINS-NAcb and aINS-BLA neurons (identified by “opto-tagging,” light-based stimulation to identify particular cells). We predict that greater aINS firing will correlate with higher front-loading, especially aINS-BLA in HAD and aINS-NAcore in P rat. Before alcohol, we also examine aINS activity during anxiety, sedation, and sucrose exposure, to determine whether aINS encoding of these states predicts excessive intake level. Aim 3 uses projection-specific inhibition methods to specifically suppress aINS-NAcore or aINS-BLA and test the hypothesis that excessive alcohol intake is more affected by aINS-NAcore inhibition in P rats and aINS-BLA inhibition in HADs. Together, proposed studies have strong interactions and synergy with other Components, and, if successful, the positive impact of our work will be the discovery of key aINS mechanisms that drive front-loading and sustained high alcohol drinking, which would improve AUD treatment.