Isolation and expression of a mouse CB1 cannabinoid receptor gene - Comparison of binding properties with those of native CB1 receptors in mouse brain and N18TG2 neuroblastoma cells
Isolation and expression of a mouse CB1 cannabinoid receptor gene - Comparison of binding properties with those of native CB1 receptors in mouse brain and N18TG2 neuroblastoma cells
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DOI：
10.1016/s0006-2952(96)00727-7
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发表时间：
1997-01-24
影响因子：
5.8
通讯作者：
Tao, Q
中科院分区：
文献类型：
--
作者：
Abood, ME;Ditto, KE;Tao, Q
研究大麻素药理学的主要动物模型是小鼠。然而，小鼠大麻素受体（CB 1）基因的结构和功能表达尚未报道。我们克隆并表达了小鼠CB 1受体的基因，并将其特性与脑和N18 TG 2神经母细胞瘤细胞中的天然小鼠CB 1受体的特性进行了比较。从小鼠129品系基因组文库中分离到小鼠CB 1基因。小鼠CB 1基因组克隆的6-kb BamHI片段的序列分析表明，小鼠和大鼠之间的核酸同一性为95%（氨基酸同一性为99.5%），小鼠和人之间的核酸同一性为90%（氨基酸同一性为97%）。对小鼠CB 1基因组克隆的5'非翻译序列的检查显示，在翻译起始位点上游约60 bp处有一个剪接位点，表明可能存在CB 1受体的剪接变体。小鼠CB 1受体的编码区在293细胞中稳定表达，并测定了[H-3]SR 141716 A和[H-3]CP-55，940的结合。使用[H-3]SR 141716 A获得的B-max和K-d值（分别为921 +/- 58 fmol/mg和0.73 +/- 0.13 nM）与脑中天然小鼠CB 1受体的相似（B-max为1.81 +/- 0.44 pmol/mg，K-d为0.16 +/- 0.01 nM）和N18 TG 2细胞（B-max为197 +/- 29 fmol/mg，K-d为0.182 +/- 0.08 nM）。小鼠CB 1受体基因组克隆将为研究小鼠CB 1受体的功能和调控提供有用的工具。版权所有（C）1996 Elsevier Science Inc.
The predominant animal model in which the pharmacology of cannabinoids is studied is the mouse. Nonetheless, the structure and functional expression of the mouse cannabinoid receptor (CB1) gene have not been reported. We have cloned and expressed the gene for the mouse CB1 receptor and compared its properties with those of native mouse CB1 receptors in brain and N18TG2 neuroblastoma cells. The mouse CB1 gene was isolated from a mouse 129 strain genomic library. Sequence analysis of a 6-kb BamHI fragment of the mouse CB1 genomic clone indicates 95% nucleic acid identity between mouse and rat (99.5% amino acid identity) and 90% nucleic acid identity (97% amino acid identity) between mouse and human. Examination of the 5' untranslated sequence of the mouse CB1 genomic clone revealed a splice junction site approximately 60 bp upstream from the translation start site, indicating the possibility of splice variants of the CB1 receptors. The coding region of the mouse CB1 receptor was stably expressed in 293 cells, and binding by [H-3]SR 141716A and [H-3]CP-55,940 was determined. The B-max and K-d values obtained with [H-3]SR 141716A (921 +/- 58 fmol/mg and 0.73 +/- 0.13 nM, respectively) were similar to those of native mouse CB1 receptors in brain (B-max of 1.81 +/- 0.44 pmol/mg, K-d of 0.16 +/- 0.01 nM) and N18TG2 cells (B-max of 197 +/- 29 fmol/mg, K-d of 0.182 +/- 0.08 nM). The mouse CB1 receptor genomic clone will be a useful tool for studying the function and regulation of the CB1 receptor in mice. Copyright (C) 1996 Elsevier Science Inc.