Electrophysiology of alcohol in extended amygdala

描述（由申请人提供）：在INIA-West联盟下，这项竞争性U 01更新申请基于行为发现，即中央杏仁核（CeA）是应激反应和酒精依赖的关键脑区，并且这些行为涉及几种CeA递质（GABA，谷氨酸）和神经肽（CRF，阿片类药物和甘丙肽）。我们发表的CeA中这些系统的电生理学研究为拟议的新研究提供了一个切入点：明确需要对分子组分（例如，鼠疫布莱德诺夫和其他人）的INIA西部参与过量饮酒。因此，我们现在建议使用电生理和细胞化学方法来研究神经炎症因子（脂多糖{LPS}，Toll样受体4 {TLR 4}，CD 14，细胞因子）在酒精偏好和过度饮酒中的作用的假设。为了在细胞水平上验证这一假设，我们提出了4个具体的目的：1）通过LPS灌流或腹腔注射，在野生型（WT）和CD 14敲除（KO）小鼠的CeA中评估TLR 4活化在乙醇和CRF对CeA切片中GABA能和谷氨酸能传递的影响中的作用; 2）评估TLR 4致炎细胞因子IL-1 β、TNF α和IL-6对WT小鼠CeA中膜和突触测量的影响; 3）确定LPS、CRF或慢性乙醇是否增加CeA中炎症的细胞化学体征; 4）确定特定目的1-3中所见的LPS、细胞因子、乙醇或CRF对CeA的电生理或细胞化学作用是否可以通过用某些抗炎药物预处理来逆转。因此，这些拟议的研究代表了评估其他INIA West组件所建议的新兴基因靶点的细胞位点和作用机制的新步骤，以支持酒精偏好或过量饮酒，并可能进一步验证逆转或预防酒精效应和过量饮酒的药物靶点。 公共卫生相关性：这个项目将研究细胞和突触机制可能是最近发现的饮酒对大脑的炎症影响的基础。由于这种神经炎症效应也被认为会导致过度饮酒，因此目前的研究也代表了一个新的方向，试图验证用于预防或治疗过度饮酒和酒精中毒的药物靶点。

DESCRIPTION (provided by applicant): This competitive U01 renewal application, under the INIA-West consortium, is based on behavioral findings that the central amygdala (CeA) is a key brain area underlying stress reactions and alcohol dependence, and that these behaviors involve several CeA transmitters (GABA, glutamate) and neuropeptides (CRF, opioids and galanin). Our published electrophysiological studies of these systems in CeA provide an entry point for proposed new studies: a stated need to pursue physiological evaluation of the function of gene products suggested by the molecular components (e.g., of Y. Blednov and others) of the INIA-West to be involved in excessive alcohol drinking. Therefore, we now propose to use electrophysiological and cytochemical methods to investigate the hypothesis of a role for neuroinflammatory factors (lipopolysaccharide {LPS}, toll-like receptor 4 {TLR4}, CD14, cytokines) in alcohol preference and excessive drinking. To test this hypothesis at the cellular level, we propose 4 Specific Aims: 1) To assess the role of TLR4 activation in effects of ethanol and CRF on GABAergic and glutamatergic transmission in CeA slices by LPS superfusion or i.p. injection in CeA of wild type (WT) and CD14 knockout (KO) mice; 2) To assess effects of the TLR4-g en e rated inflammatory cytokines IL-ip, TNFa, and IL-6 on membrane and synaptic measures in CeA of WT mice; 3) To determine if the LPS, CRF or chronic ethanol increase cytochemical signs of inflammation in CeA; 4) To determine if the electrophysiological or cytochemical effects of LPS, cytokines, ethanol or CRF on CeA seen in Specific Aims 1-3 can be reversed by pre-treatment with certain anti-inflammatory drugs. These proposed studies thus represent new steps toward evaluating the cellular sites and mechanisms of action of the emerging gene targets suggested by other INIA West components to underlie alcohol preference or excessive drinking, and may further validate drug targets for reversal or prevention of alcohol effects and excessive drinking. PUBLIC HEALTH RELEVANCE: This project will examine the cellular and synaptic mechanisms likely to underlie the recently discovered inflammatory effects on the brain of alcohol drinking. Because such neuro-inflammatory effects are also suggested to lead to excessive drinking, the present studies also represent a new direction in attempts to validate drug targets for the prevention or treatment of excessive drinking and alcoholism.