The role of central amygdala somatostatin signaling in binge ethanol drinking

项目摘要 酗酒是最常见的酒精摄入形式，占美国酒精摄入量的四分之三以上。 与美国酒精使用相关的经济负担。除了相关的负面健康影响 如心脏病和糖尿病，暴饮暴食是酒精发展的一个重要危险因素。 依赖性是一个普遍的公共卫生问题。尽管如此，可用的治疗方案有限 酗酒或酒精使用障碍。中央杏仁核内的应激相关神经肽 (CeA)已成为新型药物疗法开发的重要目标，然而其中一个的作用 大脑中表达最密集的“抗压力”神经肽，生长抑素（SST），在酗酒中 行为基本上被忽视了。这一遗漏特别值得注意，因为SST是众所周知的， 神经调节剂和神经营养因子，使其能够通过与神经元的直接相互作用改变CeA活性， 通过调节星形胶质细胞的活性来间接调节。我假设酒精摄入过量会使SST失调 受体表达的神经和星形胶质细胞群体，进一步，星形胶质细胞的调节， 对SST表达细胞的抑制性传递同样会失调，导致表达能力降低。 这种重要的“抗应激”肽调节CeA活性。这些失调将完全准备好 显着有助于众所周知的增加CeA活动发生在酒精 依赖解剖酒精诱导的非肿瘤细胞中CeA SST系统表达和功能的改变 因此，依赖性动物将能够发现针对这种危险摄入模式的新疗法， 提供了重要的见解，初步乙醇诱导的变化的一个区域，关键参与发展 和酒精依赖的维持。 这些建议的研究的K 00阶段采用1）星形胶质细胞形态学的尖端分析（使用 免疫组织化学和共聚焦显微镜技术的培训），以表征乙醇诱导的变化 SST受体在CeA星形胶质细胞和神经元群体中的表达，以及总体 星形胶质细胞形态和星形胶质细胞突触周鞘在含SST的突触上的表达 神经元;和2）使用离体全细胞膜片钳电生理学来评估GABA能神经元的改变。 传递到表达SST的细胞，以及星形胶质细胞对这种传递的调节变化（使用训练 在双重修补技术中沉默星形胶质细胞活性）。拟议研究的R 00阶段将使用这些 结合我在化学遗传学技术和行为药理学方面的现有专业知识， 1）评估表达SST的CeA神经元对终纹床核（BNST）投射的作用 2）评估酗酒引起的SST受体表达的变化， BNST和BNST星形胶质细胞对BNST内CeA SST末端活性的调节。

PROJECT SUMMARY Binge ethanol consumption is the most common form of alcohol intake and represents over three-quarters of the financial burden associated with alcohol use in the United States. In addition to associated negative health effects such as heart disease and diabetes, binge intake is a significant risk factor for the development of alcohol dependence, is a widespread public health concern. Despite this, there are limited treatment options available for binge drinking or alcohol use disorders as a whole. Stress-related neuropeptides in the central amygdala (CeA) have emerged as important targets for development of novel pharmacotherapies, however the role of one of the most densely expressed ‘anti-stress’ neuropeptides within the brain, somatostatin (SST), in binge drinking behavior has gone essentially neglected. This omission is of particular note as SST is well known as both a neuromodulator and a neurotrophic factor, allowing it to alter CeA activity by direct interactions with neurons and indirectly through modulating astrocytic activity. I hypothesize binge-ethanol intake will dysregulate SST receptors expressed on both neural and astrocytic populations and, further, that astrocyte modulation of inhibitory transmission on to SST-expressing cells will likewise be dysregulated, resulting in decreased ability of this important ‘anti-stress’ peptide to regulate CeA activity. These dysregulations would be perfectly poised to significantly contribute to the well-established increase in CeA activity known to occur during alcohol dependence. Dissecting binge-ethanol induced alterations in CeA SST system expression and function in non- dependent animals will therefore enable both discovery of novel treatments for this risky pattern of intake and provide important insights into initial ethanol-induced alterations of a region critically involved in the development and maintenance of alcohol dependence. The K00 phase of these proposed studies employ 1) cutting-edged analysis of astrocyte morphology (using training in immunohistochemical and confocal microscopy techniques) to characterize ethanol-induced changes in SST receptor expression on astrocytic and neuronal populations in the CeA, as well as changes in overall astrocyte morphology and expression of the astroglial perisynaptic sheath at synapses onto SST-containing neurons; and 2) use of ex vivo whole cell patch-clamp electrophysiology to assess alterations in GABAergic transmission on to SST-expressing cells, and changes in astrocyte modulation of this transmission (using training in duel-patching technique to silence astrocyte activity). The R00 phase of the proposed studies will use these techniques in combination with my existing expertise in chemogenetic technology and behavioral pharmacology to 1) assess the role of the SST-expressing CeA neuron to bed nucleus of the stria terminalis (BNST) projection in binge-drinking behavior; and 2) assess binge-ethanol induced changes in SST receptor expression within the BNST and BNST astrocyte modulation of CeA SST terminal activity within the BNST.