title: 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.
abstract: 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 [3H]SR 141716A and [3H]CP-55,940 was determined. The Bmax and Kd values obtained with [3H]SR 141716A (921 +/- 58 fmol/mg and 0.73 +/- 0.13 nM, respectively) were similar to those of native mouse CB1 receptors in brain (Bmax of 1.81 +/- 0.44 pmol/mg, Kd of 0.16 +/- 0.01 nM) and N18TG2 cells (Bmax of 197 +/- 29 fmol/mg, Kd 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.
authors.affiliation: Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond 23298, USA. mabood@gems.vcu.edu
authors.forename: M E
authors.initials: ME
authors.lastname: Abood
authors.forename: K E
authors.initials: KE
authors.lastname: Ditto
authors.forename: M A
authors.initials: MA
authors.lastname: Noel
authors.forename: V M
authors.initials: VM
authors.lastname: Showalter
authors.forename: Q
authors.initials: Q
authors.lastname: Tao
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000378:metabolism; D010880:Piperidines / Q000378:metabolism; D011720:Pyrazoles / Q000378:metabolism; D051381:Rats; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000077285:Rimonabant; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: D010880:Piperidines; D011720:Pyrazoles; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug; D000077285:Rimonabant