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Fish Physiology and Biochemistry

, Volume 39, Issue 5, pp 1287–1296 | Cite as

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

  • Erika Cottone
  • Valentina Pomatto
  • Fulvio Cerri
  • Ezio Campantico
  • Ken Mackie
  • Massimiliano Delpero
  • Alda Guastalla
  • Claudio Dati
  • Patrizia Bovolin
  • Maria Fosca Franzoni
Article

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.

Keywords

Endocannabinoid system Carassius auratus Sequence identity Western-blotting qReal-Time PCR 

Notes

Acknowledgments

The authors wish to thank Dr. Marco Del Giudice and Dr. Sergio Castellano for their suggestions about the statistical analysis. This research was financially supported by the Italian Ministry of University Research and Technology (PRIN 2005, PRIN 2008 and ex-60 % 2008 to MFF).

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Erika Cottone
    • 1
  • Valentina Pomatto
    • 1
  • Fulvio Cerri
    • 1
  • Ezio Campantico
    • 1
  • Ken Mackie
    • 2
  • Massimiliano Delpero
    • 1
  • Alda Guastalla
    • 1
  • Claudio Dati
    • 1
  • Patrizia Bovolin
    • 1
  • Maria Fosca Franzoni
    • 1
  1. 1.Department of Life Science and Systems BiologyUniversity of TurinTurinItaly
  2. 2.Gill Center for Biomolecular Science, Department of Psychological and Brain SciencesIndiana UniversityBloomingtonUSA

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