Electrophysiology of alcohol in extended amygdala

描述（由申请人提供）：本修订申请补充资金是对通知号（NOT-OD-09-058）和通知标题的回应：NIH宣布为竞争性修订申请提供恢复法案资金。该应用程序的父代INIA West U01基于行为研究发现，中央杏仁核（CeA）是应激反应和滥用药物强化特性的关键脑区，这些行为涉及几种CeA递质（GABA，谷氨酸）和神经肽（CRF，阿片类药物和丙氨酸）。我们假设CeA中的这些神经化学系统与依赖动物过量饮酒有关。我们对这些系统在CeA神经元中的电生理研究要么已经发表（CRF1， mu和delta阿片受体的作用），要么正在进行（kappa阿片受体和丙氨酸受体的作用）。然而，由于最近表明INIA West需要开始对INIA West的分子成分所提示的基因产物的功能进行生理评估，从而参与过量饮酒，我们提出了这一应用的新研究。具体来说，我们将使用电生理方法来研究Y. Blednov Research U01发现的炎症因子（脂多糖{LPS}， TLR4, CD14，细胞因子）在CeA中酒精相关的神经元功能中的细胞作用。为此，我们将采用体外脑切片电生理记录方法，研究LPS、乙醇和CRF对野生型和CD14敲除小鼠CeA神经元中gaba能和谷氨酸能传递的相互作用。因此，这些提出的新研究代表了评估其他INIA West成分提示的导致酒精偏好或过度饮酒的新兴基因靶点的细胞位置和作用机制的新的早期步骤。此外，通过这个申请，我们也试图保留我们的高级博士后研究员的工作，并允许我们的高级技术人员全力以赴。

DESCRIPTION (provided by applicant): This revision application for supplemental funds is in response to Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. The parent INIA West U01 from which this application is derived is based on behavioral findings that the central amygdala nucleus (CeA) is a key brain area underlying stress reactions and the reinforcing properties of abused drugs, and that these behaviors involve several CeA transmitters (GABA, glutamate) and neuropeptides (CRF, opioids and galanin). We hypothesized that these neurochemical systems in the CeA are involved in the excessive ethanol drinking seen in dependent animals. Our electrophysiological studies of these systems in CeA neurons are either published (the role of CRF1, mu and delta opioid receptors) or ongoing (role of kappa opioid and galanin receptors). However, due to the recently stated need of the INIA West to begin physiological evaluation of the function of gene products suggested by the molecular components of the INIA West to be involved in excessive alcohol drinking, we propose the new studies of this application. Specifically, we will use electrophysiological methods to investigate the cellular role of the inflammatory factors identified by the Y. Blednov Research U01 (lipopolysaccharide {LPS}, TLR4, CD14, cytokines) in alcohol-related aspects of neuronal function in the CeA. Toward this aim we will use electrophysiological recording methods in brain slices in vitro and examine the interactions of LPS, ethanol and CRF on GABAergic and glutamatergic transmission in CeA neurons of wild type and CD14 knockout mice. These proposed new studies thus represent the new early 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. In addition, with this application we also are attempting to retain the job of our senior postdoctoral researcher and to allow full effort from our senior technician.