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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
- Publication Year :
- 2013
-
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 pufferfish 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 kDa and 40 kDa, and other faint bands with apparent molecular masses around 70 kDa, 57 kDa 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 quantitative Real-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 non-mammalian vertebrates.
- Subjects :
- Cannabinoid receptor
endocannabinoid system
carassius auratus
sequence identity
western-blotting
qReal-Time PCR
Physiology
Blotting, Western
Molecular Sequence Data
Danio
Sequence Homology
Aquatic Science
Molecular cloning
Real-Time Polymerase Chain Reaction
Biochemistry
Article
Receptor, Cannabinoid, CB2
Receptor, Cannabinoid, CB1
Species Specificity
Goldfish
Cannabinoid receptor type 2
Animals
Cluster Analysis
RNA, Messenger
Cloning, Molecular
Receptor
Zebrafish
Phylogeny
DNA Primers
Base Sequence
biology
Gene Expression Profiling
Sequence Analysis, DNA
General Medicine
biology.organism_classification
Molecular biology
Endocannabinoid system
Cell biology
Gene expression profiling
nervous system
lipids (amino acids, peptides, and proteins)
sense organs
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....b16d29fe1f3ba2fe51c3a33c85ddccf4