Hypothalamic oxytocin influence on extended amygdala CRF neurons in alcohol dependence

项目摘要/摘要 酒精中毒是一种慢性复发性精神障碍，其特征是强迫性寻求和饮酒， 从娱乐用途过渡到滥用和依赖的结果。大多数酗酒者不接受治疗， 目前的药物并不适用于所有患者，这突显了新疗法的必要性。酒精 依赖导致大脑应激系统活动增强，导致消极情绪状态。 与戒断有关。神经肽催产素(OT)具有抗应激作用，全身应用催产素 减少酗酒者戒断症状的严重程度和饮酒。中央杏仁核(CEA)和床核 终纹(BNST)是大脑的两个区域，被认为是压力处理的中心，而 这些大脑区域中的促应激和抗应激神经肽对酒精依赖的发展至关重要。 CEA和BNST的突触活动对急性酒精敏感，并在行为行为中起关键作用。 乙醇消费的影响。CEA和BNST富含神经肽及其受体，包括 促肾上腺皮质激素释放因子(CRF)和催产素，以及乙醇对这些区域突触信号的影响可能是 受神经肽活动的调节。CRF与酒精引起的高度压力和焦虑有关 依赖和戒断，阻断CEA和BNST中CRF的活性可以减少饮酒。因此， 在转变为酒精依赖的过程中，抗应激和促进应激信号之间的平衡可能会被扰乱， 以过度活跃的慢性肾衰系统为特征。下丘脑室旁核和视上核的催产素神经元 下丘脑投射到CEA和BNST，投射到含有CRF神经元的特定亚区。因此， OT可能直接作用于CEA和BNST的CRF神经元，减轻戒断和饮酒的严重程度。 本项目将表征下丘脑OT神经元对CEA和BNST的CRF神经元的输入，无论是 这些回路受到酒精依赖的干扰，而这些回路又参与酒精依赖 诱导性饮酒。病毒载体介导的荧光标记物和设计型受体的特异性表达 由特制药物(DREADD)激活将允许电生理和分子表征 CEA和BNST中的OT回路，以及OT回路参与酒精依赖的行为测试 诱导性饮酒。实验研究将在K99阶段开始，并将在 在首席调查员(PI)独立实验室的一个新机构的R00阶段。朝向这份事业 目标是，在K99阶段，PI将在指导团队下进行新技术培训，包括使用 基于病毒载体的蛋白表达和突触追踪、免疫组织化学、原位杂交和 行为药理学，以补充PI的电生理学经验。在R00阶段， 指导团队将帮助在PI的独立实验室建立这些技术，以确保连续性和 研究计划的重复性。总而言之，这项工作将深入了解OT作为一种潜在的 治疗酒精和压力/焦虑症。

PROJECT SUMMARY/ABSTRACT Alcoholism is a chronic relapsing disorder characterized by compulsive seeking and consumption of alcohol, the result of a transition from recreational use to abuse and dependence. Most alcoholics do not receive treatment, and current medications do not work for all sufferers, highlighting the need for new therapeutics. Alcohol dependence induces heightened activity of brain stress systems, resulting in the negative affective state associated with withdrawal. The neuropeptide oxytocin (OT) is anti-stress, and systemic administration of OT decreases withdrawal symptom severity and drinking in alcoholics. The central amygdala (CeA) and bed nucleus of the stria terminalis (BNST) are two brain regions considered to be hubs for stress processing, and the role of pro- and anti-stress neuropeptides in these brain regions are critical for the development of alcohol dependence. Synaptic activity in the CeA and BNST is sensitive to acute alcohol, and plays a critical role in the behavioral effects of ethanol consumption. The CeA and BNST are rich in neuropeptides and their receptors, including corticotropin releasing factor (CRF) and OT, and ethanol’s effects on synaptic signaling in these regions may be modulated by neuropeptide activity. CRF is involved in the heightened stress and anxiety associated with alcohol dependence and withdrawal, and blocking CRF activity in the CeA and BNST can reduce alcohol drinking. Thus, the balance between anti- and pro-stress signaling is likely perturbed during the transition to alcohol dependence, characterized by an overactive CRF system. OT producing neurons in the paraventricular and supraoptic nuclei of the hypothalamus project to both the CeA and BNST, to specific subdivisions that contain CRF neurons. Thus, OT may act directly on CRF neurons of the CeA and BNST to decrease withdrawal severity and alcohol drinking. This project will characterize hypothalamic OT neuronal input to CRF neurons of the CeA and BNST, whether these circuits are disrupted by alcohol dependence, and involvement of these circuits in alcohol dependence induced drinking. Viral vector mediated expression of fluorescent markers and Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) will allow for electrophysiological and molecular characterization of OT circuits in the CeA and BNST, and behavioral testing of OT circuit involvement in alcohol dependence induced alcohol drinking. Experimental studies will begin during the K99 phase, and will be completed during the R00 phase at a new institution in the principal investigator’s (PI) independent laboratory. Towards this career goal, during the K99 phase, the PI will train under the mentorship team in new techniques including the use of viral vector based protein expression and synaptic tracing, immunohistochemistry, in situ hybridization, and behavioral pharmacology to complement the PI’s experience with electrophysiology. During the R00 phase, the mentorship team will help establish these techniques in the PI’s independent laboratory to ensure continuity and reproducibility of the research plan. Collectively, this work will provide insight into the role of OT as a potential therapeutic in treating alcohol and stress/anxiety disorders.