Alcohol-induced changes in stress-related neuropeptide circuitry

摘要 酒精使用障碍(AUD)是一种严重的疾病，困扰着大约1700万美国人和 每年给美国造成4000亿美元的损失。而男性被确诊的频率是女性的两倍，而 女性容易受到酒精不良影响的影响。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还没有完全被理解。动物 模型显示，雄性啮齿类动物的饮酒量从基线水平上升，而基线饮酒量更高 在雌性啮齿动物身上很明显。戒酒会使男性产生更大的焦虑 与女性相比。需要对性别差异背后的神经回路进行研究，以推动这一领域的发展 酒精神经生物学。AUD的发育涉及中央核内关键神经肽的作用 杏仁核(CEA)，包括促肾上腺皮质激素释放因子(CRF)和神经肽Y(NPY)。CRF是一种 协调应激反应的兴奋性应激神经肽，而NPY可能作为抑制性或 兴奋性神经肽。CEA广泛接受NPY能传入，富含CRF神经元。 酒精暴露和戒断使CEA中NPY信号减少，而酒精戒断增加CRF CEA中的水平。我们已经证明，CEA中的CRF神经元投射到蓝斑(LC)，蓝斑是主要的 去甲肾上腺素(NE)的前脑来源。CRF CEA-LC通路传递与情绪有关的信息。 我们认为特定的NPY CRF-CEA LC通路在饮酒和饮酒的性别差异中起中介作用 戒断后出现焦虑样行为。这项拟议工作的目标是检验总体假设 NPY、CRF-CEA、LC-NE通路在饮酒和焦虑症中的性别差异 戒断时的行为。我们将使用一个成熟的啮齿动物酒精模型，间歇性酒精 饮酒(IED)范式。在这个模型中，女性最初比男性消费更多的酒精，而男性的饮酒量增加 摄入，导致男性和女性在饮酒30天后的摄入量相匹配。我们将使用IED进行测试 酒精对饮酒和类焦虑行为影响的电路和分子机制。我们 将整合药理学、化学遗传学、行为学和细胞学方法，包括 免疫组织化学、免疫电子显微镜和示踪。Aim 1测试CRF- 包含在CEA投射到LC-NE系统中与NPY接触的神经元，以及从慢性 酒精暴露改变雄性和雌性大鼠NPY-CRF突触组织。《目标2》检验了这一假设 投射到LC的CRF-CEA神经元内的Y1r激动剂将逆转饮酒和饮酒的增加。 雄性和雌性大鼠戒酒时的焦虑样行为。从性别差异的角度来阐明 酒精戒断时CEA中影响NE传递的多肽能相互作用及NPY的作用 在通过杏仁核CRF回路调节LC-NE中，CRF在发育过程中具有重要的临床意义 AUDS的个体化治疗和新靶点。

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.