ANANDAMIDE/THC INDUCED ARACHIDONIC ACID RELEASE

本应用程序中提出的实验的目标是确定 导致大麻素诱导花生四烯酸的生化事件 放手。将进行的研究将集中在信号上 大麻素与其受体结合后的转导过程。这个 受体在这一过程中的调解问题将由 使用反义寡核苷酸除了更多 传统的配基结合方法。寡头将被设计成减少， 或者完全消除大麻受体基因的表达 并与随后花生四烯酸释放的变化进行比较。这一术语 大麻素是指包括两种外源物质，如THC， 以及内源性分子，如花生胺。将用于这些项目的系统 调查将分离、完好的小鼠腹膜细胞等 巨噬细胞、神经母细胞瘤和中国仓鼠卵巢系。这个 实验的目的是确定磷脂酶D，蛋白质的作用 激酶C、MAP激酶和磷脂酶A2在大麻素受体中的作用 刺激花生四烯酸的释放和随后产生的 二十烷类化合物。酶活性的直接测量将通过 大麻素浓度范围及与花生四烯酸的比较 在相同的条件下释放。还将进行时间课程研究 以帮助澄清所涉及的事件的顺序。特异性的抑制物 在可能的情况下，将使用各种酶来补充数据 通过直接测量获得的。 二十烷类化合物，如前列腺素E2，已被证明是重要的 在人类和动物模型中大麻素作用的介体 对它们的水平如何升高的详细了解是 重要性。最近发现了一种内源性大麻素，阿南达胺， 这可能会起到脑神经调节的作用，甚至更大 对这项提案的目标的意义，因为它的结构 自然界，即花生四烯酸的乙醇醚衍生物。这个 阿南达胺的发现对“前列腺素假说”的意义 目前还不清楚大麻素的作用，然而，它不太可能 阿南达胺的二十烷基苯丙胺的性质只是巧合。更好的 了解THC/ANANDIAME的生化作用可以提高我们的 对记忆、运动协调认知等过程的理解 能力、时间感和自我知觉。此外，一些洞察力可能会 被纳入大麻滥用的分子基础的问题。这个 最近关于大麻使用量增加的报告，特别是在 再次将注意力集中在努力理解 这种药物的作用。

The goal of the experiments proposed in this application is to determine the biochemical events that lead to cannabinoid-induced arachidonic acid release. The studies to be carried out will be focused on the signal transduction processes following cannabinoid binding to its receptor. The question of receptor mediation in this process will be addressed by the use of anti sense oligodeoxynucleotides in addition to the more conventional ligand binding approach. Oligos will be designed to reduce, or eliminate completely, the expression of the cannabinoid receptor genes and compared with subsequent changes in arachidonic acid release. The term cannabinoid is meant to include both exogenous substances, such as THC, and endogenous molecules like anandamide. The systems to be used for these investigations will be isolated, intact cells such as mouse peritoneal macrophages, neuroblastoma and Chinese-hamster ovary lines. The experiments are aimed at defining roles for phospholipase D, protein kinase C, MAP kinase and phospholipase A2 in mediating cannabinoid- stimulated release of arachidonic acid and the subsequent production of eicosanoids. Direct measurement of enzyme activities will be made over a range of cannabinoid concentrations and compared with arachidonic acid release under identical conditions. Time course studies will also be done to help elucidate the sequence of events involved. Specific inhibitors of the various enzymes will be used where possible to supplement the data obtained by direct measurement. Eicosanoids, such as prostaglandin E2, have been shown to be important mediators of cannabinoid action in humans as well as in animal models so that a detailed understanding of how their levels are elevated is of importance. The recent discovery of an endogenous cannabinoid, anandamide, that may function as a brain neuroregulator, lends even greater significance to the aims of this proposal because of its structural nature, namely, the ethanolantide derivative of arachidonic acid. The significance of the anandamide discovery to the "prostaglandin hypothesis" of cannabinoid action is not clear at this time, however, it is unlikely that the eicosanoid nature of anandamide is only coincidental. A better understanding of THC/anandamide biochemical actions could improve our understanding of processes such as memory, motor coordination cognitive ability, time sense and self-perception. In addition, some insight might be gained into the question of the molecular basis for cannabis abuse. The recent reports on the increasing use of marihuana, especially among preadolescents, has again focussed attention on efforts to understand the actions of this drug.