Role of Opiates in Alcohol-Induced Neurotoxicity- Diversity supplement

与这项资助有关的研究已经朝着确定小胶质细胞在调节乙醇中的作用的方向发展。 参与下丘脑垂体调节的R-内啡肽神经元发育过程中的细胞凋亡作用 肾上腺轴(HPA)功能。我们在过去获奖资助期内所做的工作 证明在生命早期接触酒精会导致小胶质细胞的激活和释放 影响下丘脑神经元生理功能的细胞因子和趋化因子。此外， 生命早期接触乙醇可能会使小胶质细胞群产生更多炎症 成年后应激挑战后的细胞因子。此外，调节HPA的B-内啡肽神经元 AXIS不仅是炎性细胞因子和趋化因子的靶标，也是小胶质细胞的调节剂 功能。由于缺乏信息，小胶质细胞和BEP如何相互作用来控制应激反应，以及 从大鼠神经干细胞培养纯B-内啡肽神经元的方法的可用性，我们建议 检验以下假设：i)酒精暴露刺激小胶质细胞产生炎性细胞因子 包括激活核因子-KB(NFKB)途径的肿瘤坏死因子-α和NADPH氧化酶，以诱导 发育中的B-内啡肽神经元的凋亡信号；II)锂-内啡肽神经元影响乙醇的 通过改变小胶质细胞的阿片受体功能来增加炎性细胞因子产生的能力； 以及三)早年酒精暴露使小胶质细胞群产生更多炎症 成年期压力挑战后的细胞因子。我们将采用各种最先进的技术 涉及体外分化纯B-内啡肽细胞、小胶质细胞原代培养、 酒精喂养新生儿模型，基因芯片和蛋白质组学方法，蛋白质印迹，实时荧光定量聚合酶链式反应， MeDIP检测，双重免疫细胞化学，基因敲除。我们还将在体外和体内使用 接近了。我们预计，这些研究将确定两国之间的双向沟通 下丘脑中的小胶质细胞和B-内啡肽可能预测应激异常的风险 发育期酒精暴露。 相关性(请参阅说明)： 拟议的一系列研究应继续产生有价值的数据，以便更好地了解PF 乙醇对发育中的B-内啡肽神经元的神经毒性作用。此外，拟议的研究应 识别作用于新靶点的化合物以抑制广泛的促炎因子释放 来自过度激活的小胶质细胞，这可能是防止B-内啡肽神经元死亡的关键。

Research pertaining to this grant has evolved toward determining the role of microglia in mediating ethanol apoptotic action on developing R-endorphin neurons involved in regulation ofthe hypothalamic pituitary adrenal axis (HPA) functions. The work conducted by us during the past funding period of the MERIT award demonstrated that alcohol exposure during early life causes activation of microglial cells and releases cytokines and chemokines that impact the physiological functions of hypothalamic neurons. Furthermore, early life exposure of ethanol may program the microglial cell population to produce more inflammatory cytokines following a stress challenge in the adulthood. Also, B-endorphin neurons that regulate the HPA axis are not only a target of inflammatory cytokines and chemokines but also a modulator of microglial cell functions. Given the lack of information how microglia and BEP interact to control stress responses and the availability of methods of culturing pure B-endorphin neurons from rat neuronal stem cells, we propose to test the following hypotheses: i) alcohol exposures stimulate microglia to produce inflammatory cytokines including TNF-a that activates the nuclear factor-KB (NFKB) pathway and NADPH oxidase to induce apoptotic signaling in developing B-endorphin neurons; ii) li-endorphin neurons influences the ethanol's ability to increase inflammatory cytokines production by altering opioid receptor functions in microglial cells; and iii) early-life alcohol exposures program the microglial cell population to produce more inflammatory cytokines following a stress challenge during the adulthood. We will employ various state-of-the art techniques involving in vitro differentiated pure B-endorphin cells, primary cultures of microglial cells, neonatal model of alcohol feeding, microarray and proteomic approaches, western blot, realtime PCR, MeDIP assay, double immunocytochemistry, gene knocking-down. We will also use both in vitro and in vivo approaches. We anticipate these studies will identify whther a bi-directional communication between microglia and B-endorphin within the hypothalamus may predict risk for stress abnormality in a model of developmental alcohol exposures. RELEVANCE (See instructions): The proposed series of studies should continue to generate valuable data for better understanding pf ethanol's neurotoxic action on developing B-endorphin neurons. Additionally, the proposed research should identify compounds acting on novel targets to inhibit the release of a wide range of proinflammatory factprs from overactivated microglia that might be critical for preventing of B-endorphin neuronal death.