Neurological Diseases Research, Global Pharmaceutical Research & Development, Abbott Laboratories, R47W, AP9A, 100 Abbott Park Road, Abbott Park, IL 60064, USA.
Eur J Pharmacol. 2011 Jun 1;659(2-3):161-8. doi: 10.1016/j.ejphar.2011.03.020. Epub 2011 Mar 31.
The behavioral effects evoked by cannabinoids are primarily mediated by the CB(1) and CB(2) cannabinoid receptor subtypes. In vitro pharmacology of cannabinoid receptors has been elucidated using recombinant expression systems expressing either CB(1) or CB(2) receptors, with limited characterization in native cell lines endogenously expressing both CB(1) and CB(2) receptors. In the current study, we report the molecular and pharmacological characterization of the F-11 cell line, a hybridoma of rat dorsal root ganglion neurons and mouse neuroblastoma (N18TG2) cells, reported to endogenously express both cannabinoid receptors. The present study revealed that both receptors are of mouse origin in F-11 cells, and describes the relative gene expression levels between the two receptors. Pharmacological characterization of the F-11 cell line using cannabinoid agonists and antagonists indicated that the functional responses to these cannabinoid ligands are mainly mediated by CB(1) receptors. The non-selective cannabinoid ligands CP 55,940 and WIN 55212-2 are potent agonists and their efficacies in adenylate cyclase and MAPK assays are inhibited by the CB(1) selective antagonist SR141716A (SR1), but not by the CB(2) selective antagonist SR144528 (SR2). The endocannabinoid ligand 2AG, although not active in adenylate cyclase assays, was a potent activator of MAPK signaling in F-11 cells. The analysis of CB(1) and CB(2) receptor gene expression and the characterization of cannabinoid receptor pharmacology in the F-11 cell line demonstrate that it can be used as a tool for interrogating the endogenous signal transduction of cannabinoid receptor subtypes.
大麻素引起的行为效应主要由 CB(1) 和 CB(2) 大麻素受体亚型介导。使用表达 CB(1) 或 CB(2) 受体的重组表达系统阐明了大麻素受体的体外药理学，而在天然细胞系中对同时表达 CB(1) 和 CB(2) 受体的受体进行了有限的表征。在本研究中，我们报告了 F-11 细胞系的分子和药理学特征，该细胞系是大鼠背根神经节神经元和小鼠神经母细胞瘤（N18TG2）细胞的杂交瘤，据报道该细胞系内源性表达两种大麻素受体。本研究表明，F-11 细胞中的两种受体均源自小鼠，并描述了两种受体之间的相对基因表达水平。使用大麻素激动剂和拮抗剂对 F-11 细胞系进行的药理学特征分析表明，这些大麻素配体的功能反应主要由 CB(1) 受体介导。非选择性大麻素配体 CP 55,940 和 WIN 55212-2 是强效激动剂，其在腺苷酸环化酶和 MAPK 测定中的效力被 CB(1) 选择性拮抗剂 SR141716A（SR1）抑制，但不受 CB(2) 选择性拮抗剂 SR144528（SR2）抑制。内源性大麻素配体 2AG 虽然在腺苷酸环化酶测定中不活跃，但在 F-11 细胞中是 MAPK 信号转导的有效激活剂。F-11 细胞中 CB(1) 和 CB(2) 受体基因表达的分析和大麻素受体药理学的表征表明，它可作为研究大麻素受体亚型内源性信号转导的工具。
