Project 1

该项目的目标是解决有关内源性大麻素系统在疼痛，奖励和药物依赖中的作用的几个悬而未决的问题。有强有力的证据表明，两种主要的内源性大麻素，大麻素（AEA）和2-花生四烯酸甘油（2-AG）在这些病理状态中发挥相当大的影响，但它是极不可能的，他们只是扮演多余的角色。因此，本项目的目标之一是描述它们在这些病理条件下的作用。内源性大麻素的合成和代谢途径尚未完全了解，这一事实带来了挑战 在体内条件下系统地操纵它们的水平。因此，一个主要的目标是开发有效的和选择性的酶抑制剂，可用于操纵内源性大麻素在体内。AEA和2-AG的合成和代谢酶抑制剂将由Razdan博士制备（项目2）。Cravatt博士（项目3）确定的酶选择性将由我们在大麻素活性的行为测试中进行评估。第二种方法是建立小鼠的表型， 内源性大麻素合成和代谢酶产生的克拉瓦特博士。同时，我们清楚地知道，结构上与AEA和2-AG相关的几类脂质通过影响合成和代谢途径、直接作用于大麻素受体或作用于CB{1}受体位点上新发现的变构位点来影响内源性大麻素系统。为了解决这些问题，我们将对CB{1}受体的合成变构配体[由Razdan和Mechoulam博士制备（项目4）]和Mechoulam博士提供的推定内源性大麻素进行体外和体内评价。补充博士。 Mechoulam的努力，以确定新的内源性大麻素，我们将建立在不同的实验条件下，以确定可能是内源性大麻素或相关的脂质介质的脂质在选定的大脑区域的小鼠脂质谱。我们将利用现有的以及这些新发现，进一步探索内源性大麻素系统参与疼痛，奖励和依赖。重点将放在炎症和神经性疼痛模型。我们将确定细胞因子和趋化因子参与内源性大麻素抗炎作用的程度。将使用遗传学和药理学方法研究受体的作用机制和潜在的神经底物。 同样的综合方法将用于药物辨别和进食行为，以建立操纵内源性大麻素系统的药剂的THC样特征，并建立转基因小鼠的表型行为。总的来说，这些研究将确定可能直接或间接作用于内源性大麻素系统的内源性物质，并阐明 AEA和2-AG在疼痛、奖赏和药物依赖中的作用。

The goal of this project is to address several unresolved questions regarding the role of the endocannabinoid system in pain, reward and drug dependence. There is strong evidence that the two major endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG) exert considerable influence in these pathological states, yet it is highly unlikely that they are simply playing redundant roles. Therefore, one of the objectives of this project is to delineate their roles in these pathological conditions. The fact that the synthetic and metabolic pathways for endocannabinoids are not completely understood creates a challenge in systematically manipulating their levels under in vivo conditions. Therefore, a major goal is to develop potent and selective enzyme inhibitors that can be used to manipulate endocannabinoids in vivo. Synthetic and metabolic enzyme inhibitors for AEA and 2-AG will be prepared by Dr. Razdan (Project 2). Those identified by Dr. Cravatt (Project 3) as enzyme selective will be evaluated by us in behavioral battery of tests for cannabinoid activity. The second approach is to establish the phenotypes of mice deficient in endocannabinoid synthetic and metabolic enzymes generated by Dr. Cravatt. At the same time, we are well aware that several classes of lipids structurally related to AEA and 2-AG impact the endocannabinoid system by influencing synthetic and metabolic pathways, acting directly on cannabinoid receptors, or acting at the newly discovered allosteric site on the CB{1} receptor site. To address these questions, we will conduct in vitro and in vivo evaluation of synthetic allosteric ligands for CB{1} receptors [prepared by Drs. Razdan and Mechoulam (Project 4)], and putative endocannabinoids provided by Dr. Mechoulam. To complement Dr. Mechoulam's efforts to identify new endocannabinoids, we will establish lipid profiles in selected brain regions of mice under different experimental conditions in an effort to identify lipids that may be endocannabinoids or relevant lipid mediators. We will use existing as well as these new discoveries to further explore the involvement of the endocannabinoid system in pain, reward and dependence. Emphasis will be placed on inflammatory and neuropathic pain models. We will determine the extent to which cytokines and chemokines are involved in endocannabinoid anti-inflammatory effects. The receptor mechanisms of action and underlying neural substrates will be investigated using genetic and pharmacological approaches. The same comprehensive approach will be employed in drug discrimination and feeding behavior, to establish THC-like profiles of agents that manipulate the endocannabinoid system and to establish phenotypic behavior of genetically modified mice. Collectively, these studies will identify endogenous substances that may act either directly or indirectly on the endogenous cannabinoid system, and elucidate the role that AEA and 2-AG play in pain, reward, and drug dependence.