Alcohol-induced changes in stress-related neuropeptide circuitry

摘要 酒精使用障碍（AUD）是一种严重的医疗状况，困扰着大约1700万美国人， 每年花费美国4000亿美元虽然男性被诊断出的频率是女性的两倍， 妇女容易受到酒精的不利影响。尽管在患病率和影响方面存在性别差异， 在AUD中，调节酒精使用影响的性别差异的因素尚未完全了解。动物 模型显示，在雄性啮齿动物中，饮酒量从基线增加，而基线饮酒量增加 在雌性啮齿类动物中很明显。长期酒精暴露的戒断会导致男性更大的焦虑 与女性相比。需要对性别差异背后的神经回路进行研究，以推动该领域的发展。 酒精神经生物学AUD的发展涉及中枢核团中关键神经肽的作用 包括促肾上腺皮质激素释放因子（CRF）和神经肽Y（NPY）。CRF是一个 兴奋性应激神经肽协调应激反应，而NPY可能作为抑制性或 兴奋性神经肽CeA普遍接受NPY能传入，并富含CRF神经元。 酒精暴露和酒精戒断减少了CeA中的NPY信号，而酒精戒断增加了CRF 在CeA的水平。我们已经表明，CRF神经元在CeA项目的蓝斑（LC），一个主要的 去甲肾上腺素（NE）的前脑来源。CRF CeA-LC通路传递情绪相关信息。 我们认为，特定的神经肽Y CRF-CeA LC通路介导了饮酒的性别差异， 戒断后的焦虑样行为拟议工作的目标是测试总体假设 特异性NPY CRF-CeA LC-NE通路介导了饮酒和焦虑样的性别差异。 撤退时的行为。我们将使用一个完善的啮齿动物酒精模型，即无障碍乙醇 饮酒（IED）模式。在这个模型中，女性最初比男性消费更多的酒精，而男性则逐步升级 摄入量，导致男性和女性在饮酒30天后摄入量匹配。我们会用简易爆炸装置 回路和分子机制介导酒精对饮酒和焦虑样行为的影响。我们 将整合药理学、化学遗传学、行为学和细胞学方法，包括 免疫组化、免疫电镜和束示踪。AIM 1测试了CRF- 含有神经肽Y接触的神经元投射到LC-NE系统， 酒精暴露改变了雄性和雌性大鼠的NPY-CRF突触组织。目标2检验假设 投射到LC的CRF-CeA神经元内的Y1 r激动将逆转饮酒量的增加， 雄性和雌性大鼠在酒精戒断期间的焦虑样行为。阐明性别差异， CeA中影响酒精戒断时NE传递的肽能相互作用，以及NPY的作用 通过杏仁核CRF回路调节LC-NE具有重要的临床意义， 个体化治疗和治疗AUD的新靶点。

ABSTRACT Alcohol use disorders (AUDs) are serious medical conditions that afflict approximately 17 million Americans and cost the U.S. $400 billion annually. Whereas men are diagnosed twice as frequently as women, a greater vulnerability to adverse effects of alcohol occurs in women. Despite the sex differences in prevalence and effects of AUD, the factors mediating the sex differences in the effects of alcohol use are not fully understood. Animal models showed an escalation of alcohol drinking from baseline in male rodents while higher baseline drinking was evident in female rodents. Withdrawal from chronic alcohol exposure induced a greater anxiety in male compared to female. Research is needed on the neural circuitry underlying sex differences to advance the field of alcohol neurobiology. AUD development involves the actions of critical neuropeptides in the central nucleus of the amygdala (CeA), including corticotropin-releasing factor (CRF) and neuropeptide Y (NPY). CRF is an excitatory stress neuropeptide that orchestrates stress responses, while NPY may act as an inhibitory or excitatory neuropeptide. The CeA ubiquitously receives NPY-ergic afferents and is enriched with CRF neurons. Alcohol exposure and withdrawal reduced the NPY signaling in the CeA while alcohol withdrawal increased CRF levels in the CeA. We have shown that CRF neurons in the CeA project to the locus coeruleus (LC), a major source of norepinephrine (NE) to the forebrain. The CRF CeA-LC pathway relays emotion-related information. We propose that the specific NPY CRF-CeA LC pathway mediates sex differences in alcohol drinking and anxiety-like behavior following withdrawal. The goal of the proposed work is to test the overarching hypothesis that the specific NPY CRF-CeA LC-NE pathway mediates sex differences in alcohol drinking and anxiety-like behavior during withdrawal. We will use a well-established rodent alcohol model, the intermittent-access ethanol drinking (IED) paradigm. In this model, females initially consume more alcohol than males, while males escalate intake, resulting in matched intake in males and females following 30 days of drinking. We will use IED to test circuit and molecular mechanisms mediating alcohol effects on alcohol drinking and anxiety-like behavior. We will integrate pharmacological, chemogenetics, behavioral, and cellular approaches including immunohistochemistry, immunoelectron microscopy and tract tracing. AIM 1 tests the hypothesis that CRF- containing neurons contacted by NPY in the CeA project to the LC-NE system, and that withdrawal from chronic alcohol exposure alters the NPY-CRF synaptic organizations in male and female rats. Aim 2 tests the hypothesis that Y1r agonism within the CRF-CeA neurons that project to LC will reverse the elevated alcohol drinking and anxiety-like behavior during alcohol withdrawal in male and female rats. Elucidating sex differences in the peptidergic interactions in the CeA that impact NE transmission under alcohol withdrawal, and the role of NPY in modulating the LC-NE via the amygdalar CRF circuitry has significant clinical implications in developing individualized therapies and novel targets for the treatment of AUDs.