Hypothalamic oxytocin influence on extended amygdala CRF neurons in alcohol dependence

项目总结/摘要 酒精中毒是一种慢性复发性疾病，其特征是强迫性寻求和消费酒精， 从娱乐性使用到滥用和依赖的过渡。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，这突出了对新疗法的需求。醇 依赖性会引起大脑应激系统的高度活动，从而导致消极的情感状态 与撤退有关。神经肽催产素（OT）具有抗应激作用，全身给予OT 减少戒断症状的严重程度和酗酒者的饮酒。中央杏仁核（CeA）和床核 终纹（BNST）是两个大脑区域，被认为是压力处理的枢纽， 这些脑区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，并在行为学中起着关键作用。 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OT，以及乙醇对这些区域突触信号传导的影响可能是 由神经肽活性调节。CRF与酒精相关的高度压力和焦虑有关 阻断CeA和BNST的CRF活性可减少饮酒。因此，在本发明中， 抗应激信号和促应激信号之间的平衡在向酒精依赖的转变过程中可能被扰乱， 其特点是CRF系统过于活跃。室旁核和视上核的催产素产生神经元 下丘脑投射到CeA和BNST，投射到包含CRF神经元的特定亚部。因此，在本发明中， OT可能直接作用于CeA和BNST的CRF神经元，以减轻戒断症状和饮酒。 本项目将描述下丘脑OT神经元对CeA和BNST的CRF神经元的输入， 这些回路被酒精依赖破坏，这些回路参与酒精依赖 诱导饮酒病毒载体介导的荧光标记物和设计受体的表达 设计药物激活（DREADD）将允许电生理和分子表征， CeA和BNST中的OT回路，以及酒精依赖中OT回路参与的行为测试 诱导饮酒。实验研究将在K99阶段开始，并将在 在主要研究者（PI）独立实验室的新机构进行R00阶段。为了这个职业 在K99阶段，PI将在导师团队的指导下进行新技术培训，包括使用 基于病毒载体的蛋白质表达和突触追踪，免疫组织化学，原位杂交，和 行为药理学来补充PI的电生理学经验。在R00阶段， 导师团队将帮助在PI的独立实验室建立这些技术，以确保连续性， 研究计划的可重复性。总的来说，这项工作将提供深入了解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.