title: Cannabinoid receptors are widely expressed in goldfish: molecular cloning of a CB2-like receptor and evaluation of CB1 and CB2 mRNA expression profiles in different organs.
abstract: Cannabinoids, the bioactive constituents of Cannabis sativa, and endocannabinoids, among which the most important are anandamide and 2-arachidonoylglycerol, control various biological processes by binding to specific G protein-coupled receptors, namely CB1 and CB2 cannabinoid receptors. While a vast amount of information on the mammalian endocannabinoid system does exist, few data have been reported on bony fish. In the goldfish, Carassius auratus, the CB1 receptor has been cloned and its distribution has been analyzed in the retina, brain and gonads, while CB2 had not yet been isolated. In the present paper, we cloned the goldfish CB2 receptor and show that it presents a quite high degree of amino acid identity with zebrafish Danio rerio CB2A and CB2B receptors, while the percentage of identity is lower with the puffer fish Fugu rubripes CB2, as also confirmed by the phylogenetic analysis. The sequence identity becomes much lower when comparing the goldfish and the mammalian CB2 sequences; as for other species, goldfish CB2 and CB1 amino acid sequences share moderate levels of identity. Western-blotting analysis shows the CB2 receptor as two major bands of about 53 and 40 kDa and other faint bands with apparent molecular masses around 70, 57 and 55 kDa. Since the distribution of a receptor could give information on its physiological role, we evaluated and compared CB1 and CB2 mRNA expression in different goldfish organs by means of qReal-Time PCR. Our results show that both CB1 and CB2 receptors are widely expressed in the goldfish, displaying some tissue specificities, thus opening the way for further functional studies on bony fish and other nonmammalian vertebrates.
authors.affiliation: Department of Life Science and Systems Biology, University of Turin, via Accademia Albertina 13, 10123, Turin, Italy, erika.cottone@unito.it.
authors.forename: Erika
authors.initials: E
authors.lastname: Cottone
authors.forename: Valentina
authors.initials: V
authors.lastname: Pomatto
authors.forename: Fulvio
authors.initials: F
authors.lastname: Cerri
authors.forename: Ezio
authors.initials: E
authors.lastname: Campantico
authors.forename: Ken
authors.initials: K
authors.lastname: Mackie
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Delpero
authors.forename: Alda
authors.initials: A
authors.lastname: Guastalla
authors.forename: Claudio
authors.initials: C
authors.lastname: Dati
authors.forename: Patrizia
authors.initials: P
authors.lastname: Bovolin
authors.forename: Maria Fosca
authors.initials: MF
authors.lastname: Franzoni
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D020869:Gene Expression Profiling / Q000662:veterinary; D006054:Goldfish / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D043884:Receptor, Cannabinoid, CB1 / Q000235:genetics / Q000378:metabolism*; D043885:Receptor, Cannabinoid, CB2 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D017422:Sequence Analysis, DNA / Q000662:veterinary; D017385:Sequence Homology; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D043884:Receptor, Cannabinoid, CB1; D043885:Receptor, Cannabinoid, CB2