Reuptake and hydrolysis shape endocannabinoid signaling

描述（由申请人提供）：这项工作的长期目标是确定内源性大麻素信号在新皮层中的调节机制。内源性大麻素（eCBs）从磷脂前体中分离出来，根据主细胞的需要释放，其中n -花生四烯醇乙醇酰胺（anandamide， AEA）和2-花生四烯醇甘油（2-AG）是研究最多的。然后，eCB迅速进入细胞并被水解，但失活在eCB突触传递动力学中的作用尚未被研究。脂肪酸酰胺水解酶（FAAH）在体外水解AEA和2-AG，这两种eCBs被认为是通过温度依赖性、饱和性和选择性再摄取过程重新进入细胞的。有特定的药物可以抑制FAAH和再摄取过程。本实验拟利用这些药物、FAAH敲除小鼠和全细胞膜片钳技术来研究FAAH （Aim 1）和再摄取（Aim 2）在gaba能中间神经元和皮质II/III层锥体神经元之间内源性大麻素信号传导中的作用。

DESCRIPTION (provided by applicant): The long-term objective of this work is to identify the regulatory mechanisms of endogenous cannabinoid signaling in the neocortex. The endocannabinoids (eCBs), of which N-arachidonyl ethanolamide (anandamide, AEA) and 2-arachidonyl glycerol (2-AG) are the best studied, are cleaved from phospholipid precursors and released on demand from principle cells. The eCBs are then rapidly taken into cells and hydrolyzed, but the role of inactivation in the kinetics of eCB synaptic transmission has not been investigated. Fatty acid amide hydrolase (FAAH) hydrolyzes both AEA and 2-AG in vitro, and both eCBs are thought to reenter cells through a temperature-dependent, saturable, and selective reuptake process. Specific pharmacologic agents are available to inhibit both FAAH and the reuptake process. The proposed experiments intend to make use of these agents, FAAH knockout mice, and whole-cell patch clamp techniques to investigate the role of FAAH (Aim 1) and reuptake (Aim 2) in endocannabinoid signaling between GABAergic interneurons and cortical Layer II/III pyramidal neurons.