Molecular cloning of a human cannabinoid receptor which is also expressed in testis

• DOI: 10.1042/bj2790129
• Publication Date: 2015-08-10T21:23:44Z
• Authors: C M Gérard; C Mollereau; G Vassart; M Parmentier
• Keywords: Male; Drug -- genetics; Molecular Sequence Data; Forskolin -- pharmacology; Gene Expression; Cyclohexanols -- metabolism; Cell Line; Cricetulus; Genetic; Cricetinae; Receptors; Cyclic AMP; Animals; Humans; Northern; Amino Acid Sequence; Cloning, Molecular; Receptors, Cannabinoid; Cannabinoid; Testis -- chemistry; Base Sequence; Blotting; Cannabinoids; Cannabinoids -- metabolism; Colforsin; Molecular; DNA; Sciences bio-médicales et agricoles; Blotting, Northern; Cyclohexanols; RNA -- genetics; DNA -- genetics; Brain Stem -- chemistry; RNA; Cyclic AMP -- metabolism; Transcription; Cloning; Brain Stem

A cDNA clone encoding a receptor protein which presents all the characteristics of guanine-nucleotide-binding (G-protein)-coupled was isolated from human brain stem library. The probe used (HGMP08) 600 bp DNA fragment amplified by low-stringency PCR, using genomic as template and degenerate oligonucleotide primers corresponding to conserved sequences amongst known G-protein-coupled receptors. deduced amino acid sequence encodes 472 residues shares 97.3% identity with rat cannabinoid cloned recently [Matsuda, Lolait, Brownstein, Young & Bronner (1990) Nature (London) 346, 561-564]. Abundant transcripts were detected in brain, expected, but lower amounts also found testis. same screen testis clones obtained partially sequenced, demonstrating receptors expressed both tissues. Specific binding synthetic ligand [3H]CP55940 observed on membranes Cos-7 cells transfected recombinant clone. In stably CHO-K1 cell lines, agonists mediated dose-dependent stereoselective inhibition forskolin-induced cyclic AMP accumulation. ability express mammalian should help developing more selective drugs, facilitate search for endogenous ligand(s).