Biochemistry of Anandamide, An Endogenous Cannabinoid

描述(申请人提供)：长期目标。我们的目标是了解调节大脑大麻素受体(CB1)内源性配体浓度的机制，以激活该受体。这一点很重要，因为CB1受体激活在大脑信号调节中的重要性。假设：我们假设神经元活动的变化导致内源性大麻素(ECB)从细胞内区域迅速重新分布到细胞外区域。我们进一步假设，这种重新分配是ECB从细胞内隔离位置动员、合成增加和分解代谢减少的综合结果。我们还假设ECB分解代谢的主要部位是突触前CB1受体表达的神经元。目的：我们的第一个目标是确定突触活性增加对ECB释放的调节；第二个目标是研究2-AG失活的机制；第三个目标是确定固醇载体蛋白2是否参与神经元对ECB的聚集或隔离。方法：我们将使用培养细胞和小脑切片进行这些研究。我们将使用液-质联用来测量神经元激活对ECB在组织和缓冲液之间重新分布的影响。意义：越来越清楚的是，ECB/CB1信号对在许多行为和神经精神障碍中发挥着重要作用。特别是，在动物和人类中进行的几项研究表明，改变ECB的产生可能会增强滥用药物的成瘾倾向。例如，一种CB1受体拮抗剂被认为是戒烟的一种疗法。因此，增加对CB1受体内源性激活过程的了解将有助于我们理解这种药物治疗成瘾的潜力。新颖性：我们实验室专注于使用液相色谱质谱联用来检测内源性大麻素。我们将这项技术与我们从小脑培养细胞的能力结合起来，作为实现这些目标的独特方法。

DESCRIPTION (provided by applicant): Long term goals. Our goals are to understand the mechanisms that regulate the concentrations of the endogenous ligands of the brain cannabinoid receptor (CB1) available to activate the receptor. This is important because of the importance of CB1 receptor activation in the regulation of signaling in the brain. Hypothesis: We hypothesize that changes in neuronal activity induce a rapid redistribution of the endocannabinoids (eCBs) from intracellular to extracellular domains. We further hypothesize that this redistribution occurs as a result of a combination of the mobilization of the eCBs from intracellular sequestration sites, increased synthesis and reduced catabolism. We also hypothesize that the primary sites of eCB catabolism are presynaptic, CB1 receptor expressing neurons. Aims: Our first aim is to determine regulation of eCB release in response to increased synaptic activity; the second is to study the mechanisms of inactivation of 2-AG; the third aim is to determine whether sterol carrier protein 2 is involved in the accumulation or sequestration of the eCBs by neurons. Methods: We will carry out these studies using cells in culture and cerebellar slices. We will use liquid chromatography:mass spectrometry to measure the effects of neuronal activation on the redistribution of the eCBs between tissue and buffer. Significance: It is becoming increasingly clear that the eCB/CB1 signaling pair plays important roles in many behavioral and neuropsychiatric disorders. In particular, several studies in both animals and humans suggest that altered eCB production could enhance the addictive liability of drugs of abuse. A CB1 receptor antagonist is being considered as a therapy for smoking cessation, for example. Therefore, increased understanding of the processes involved in the endogenous activation of the CB1 receptor will aid in our understanding of the potential for this drug to treat addiction. Novelty: Our laboratory has focused on the use of liquid chromatography mass spectrometry to assay endocannabinoids. We have coupled this technique with our ability to culture cells from the cerebellum as a unique approach to these goals.