Anterior Insula Projections for Alcohol Drinking/Anxiety Interactions in Female and Male Rats

摘要 酒精使用障碍（AUD）提取大量的医疗，社会和经济成本， 饮酒（CLAD），尽管有负面影响，但摄入量仍然存在，可能是一个特别的障碍， 治疗同样令人担忧的是，女性饮酒问题急剧上升，女性可能会有 更大的酒精问题。然而，对性别差异的潜在机制仍然知之甚少， 虽然焦虑样状态可能会强烈导致饮酒，但女性的饮酒率更高， 情绪障碍和AUD/焦虑共病。此外，我们的老鼠研究表明，女性有更大的 对酒精的持续反应，以及更大的焦虑样反应。因此，为了帮助更好地发展， 个性化的治疗，这是至关重要的，以了解性别差异和相似性的机制， 过度饮酒和焦虑症酒精摄入可能是由其显著的动机驱动的 前额叶（aINS）是一个重要的调节器， 伴随着情绪状态。我们发现特定的aINS投射（包括aINS-伏核， NAcb）促进男性CLAD，对仅饮酒（AOD）影响不大;一致认为，强迫性 患有AUD的女性和男性对酒精的反应激活了类似的aINS回路。然而，很少有 已知女性饮酒的特定aINS机制，或克服更高的 更普遍的挑战。此外，整体aINS抑制减少了AOD，但潜在的aINS投射（s） 完全没有被发现。在这里，我们解决的首要假设，不同的aINS电路， 驱动CLAD与AOD，特别是，特定的aINS回路介导CLAD和焦虑样行为 (ALB)在高度焦虑的个体中，这在女性中尤其有说服力。Aim 1使用特定于项目的 光遗传学抑制以检查AOD和CLAD的特定aINS投射：aINS-NAcb和aINS-NAcb。 杏仁核用于CLAD，aINS-后丘脑（pINS）用于AOD。aINS-pINS投影较大，pINS 抑制减少饮酒，但aINS-pINS几乎未在啮齿动物中研究。此外，为了更好地了解AINS 由于输出调节行为，因此确定特定aINS预测内的实际活动模式至关重要。因此，在本发明中， AIM 2使用尖端的体内电生理记录方法和“光标记”来识别击发 特定aINS预测内的模式，特别是预测女性的更高活动，这是更大的基础。 酒精的持久性最后，目标3检查了ALB与 饮酒，这些行为的aINS编码，以及特定aINS的光遗传学抑制的影响 对ALB和饮酒的预测。我们预测，高ALB大鼠将有更大的aINS编码， ALB和饮酒，以及ALB和摄入的更大光遗传学抑制，特别是在女性中。如果成功， 我们将发现驱动AOD、CLAD、ALB的关键aINS投射机制及其相互关系， 包括重要的性别差异，这可能会开启新的性别特异性AUD治疗。

Abstract Alcohol Use Disorder (AUD) extracts substantial medical, social, and economic costs, and compulsion-like alcohol drinking (CLAD), where intake persists despite negative consequences, can be a particular obstacle to treatment. Also concerning is that problem drinking in women has risen dramatically, and women can have greater alcohol problems. However, mechanisms underlying sex differences remain poorly understood, although anxiety-like states can strongly contribute to alcohol drinking, and women have greater prevalence of mood disorders and AUD/anxiety comorbidity. Also, our rat studies suggest that females have greater persistence in responding for alcohol, as well as greater anxiety-like responding. Thus, to help develop better, personalized therapies, it is essential to understand sex differences and similarities in mechanisms underlying excessive drinking and comorbidity with anxiety. Alcohol intake is likely driven by its salient motivational properties, and anterior insula (aINS) is a critical regulator of responding to important situations and regulating accompanying emotional states. We find that specific aINS projections (including aINS-Nucleus Accumbens, NAcb) promote male CLAD, with little impact on alcohol-only drinking (AOD); in agreement, compulsion-like responding for alcohol in women and men with AUD activates a similar aINS circuit. However, very little is known about specific aINS mechanisms for alcohol drinking in females, or overcoming higher challenge more generally. Also, global aINS inhibition reduces AOD, but the underlying aINS projection(s) remain completely undiscovered. Here, we address the overarching hypothesis that different aINS circuits drive CLAD vs AOD, and, especially, that particular aINS circuits mediate both CLAD and anxiety-like behavior (ALB) in higher-anxiety individuals, which is especially cogent in females. Aim 1 uses projection-specific optogenetic inhibition to examine particular aINS projections for AOD and CLAD: aINS-NAcb and aINS- amygdala for CLAD, and aINS-posterior insula (pINS) for AOD. aINS-pINS projection is large, and pINS inhibition decreases drinking, but aINS-pINS is nearly unstudied in rodent. Also, to best understand how aINS outputs regulate behavior, it is critical to identify actual activity patterns within specific aINS projections. Thus, Aim 2 uses cutting edge in vivo electrophysiology recording methods with “opto-tagging” to identify firing patterns within specific aINS projections, especially predicted higher activity in females that underlies greater persistence for alcohol. Finally, Aim 3 examines potential sex differences in the relation between ALB and alcohol drinking, aINS encoding of these behaviors, and the impact of optogenetic inhibition of specific aINS projections on both ALB and drinking. We predict that higher ALB rats will have greater aINS encoding of both ALB and drinking, and greater optogenetic inhibition of ALB and intake, particularly in females. If successful, we will discover key aINS projection mechanisms that drive AOD, CLAD, ALB, and their interrelations, including important sex differences, which may open novel, sex-specific AUD treatments.