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新皮层中内源性大麻素信号传导的调节机制。内源性大麻素（eCB），其中N-花生四烯基乙醇酰胺（anandamide，AEA）和2-花生四烯基甘油（2-AG）是最好的研究，从磷脂前体裂解，并根据需要从主细胞释放。然后eCB迅速进入细胞并水解，但失活在eCB突触传递动力学中的作用尚未研究。脂肪酸酰胺水解酶（FAAH）在体外水解AEA和2-AG，并且认为这两种eCB都通过温度依赖性、饱和性和选择性再摄取过程重新进入细胞。特定的药物可用于抑制FAAH和再摄取过程。拟议的实验打算利用这些代理商，FAAH敲除小鼠，和全细胞膜片钳技术，以调查的作用FAAH（目标1）和再摄取（目标2）的GABA能中间神经元和皮质层II/III锥体神经元之间的内源性大麻素信号。

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.