Reuptake and hydrolysis shape endocannabinoid signaling

描述(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质中的调节机制。内源性大麻素(ECB)是从磷脂前体中裂解出来并根据需要从主细胞中释放出来的，其中研究最多的是N-花生四烯基乙醇胺(AEA)和2-花生四烯基甘油(2-AG)。ECB随后被迅速带入细胞并被水解，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)在体外同时降解AEA和2-AG，这两种ECB都被认为通过依赖温度的、可饱和的和选择性的重摄取过程重新进入细胞。特定的药理制剂可用于抑制FAAH和再摄取过程。本实验拟利用FAAH基因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层锥体神经元之间的内源性大麻素信号转导中的作用。

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.