Behavioral, pharmacological, and molecular characterization of an amphibian cannabinoid receptor

Ken Soderstrom, Mark Leid, Frank L. Moore, Thomas F. Murray

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Investigation of cannabinoid pharmacology in a vertebrate with a phylogenetic history distinct from that of mammals may allow better understanding of the physiological significance of cannabinoid neurochemistry. Taricha granulosa, the roughskin newt, was used here to characterize an amphibian cannabinoid receptor. Behavioral experiments demonstrated that the cannabinoid agonist levonantradol inhibits both newt spontaneous locomotor activity and courtship clasping behavior. Inhibition of clasping was dose-dependent and potent (IC50 = 1.2 μg per animal). Radioligand binding studies using [3H]CP-55940 allowed identification of a specific binding site (K(D) = 6.5 nM, B(max) = 1,853 fmol/mg of protein) in brain membranes. Rank order of affinity of several ligands was consistent with that reported for mammalian species (K(D), nM): CP-55940 (3.8) > levonantradol (13.0) > WIN55212-2 (25.7) >> anandamide (1,665) ≃ anandamide + 100 μM phenylmethylsulfonyl fluoride (2,398). The cDNA encoding the newt CB1 cannabinoid receptor was cloned, and the corresponding mRNA of 5.9 kb was found to be highly expressed in brain. A nonclonal Chinese hamster ovary cell line stably expressing the newt CB1 cannabinoid receptor was prepared that allowed demonstration of cannabinoid-mediated inhibition of adenylate cyclase (EC 4.6.1.1) activity. This inhibition was dose-dependent and occurred at concentrations consistent with affinities determined through radioligand binding experiments. The behavioral, pharmacological, and molecular cloning results demonstrate that a CB1 cannabinoid receptor is expressed in the CNS of the roughskin newt. This amphibian CB1 is very similar in density, ligand binding affinity, ligand binding specificity, and amino acid sequence to mammalian CB1. The high degree of evolutionary conservation of cannabinoid signaling systems implies an important physiological role in vertebrate brain function.

Original languageEnglish
Pages (from-to)413-423
Number of pages11
JournalJournal of Neurochemistry
Volume75
Issue number1
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Salamandridae
Cannabinoid Receptors
Cannabinoids
Amphibians
Cannabinoid Receptor CB1
Pharmacology
Brain
Ligands
Phenylmethylsulfonyl Fluoride
Cannabinoid Receptor Agonists
Mammals
Vertebrates
Cloning
Adenylyl Cyclases
Neurochemistry
Courtship
Conservation
Animals
Demonstrations
Complementary DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Behavioral, pharmacological, and molecular characterization of an amphibian cannabinoid receptor. / Soderstrom, Ken; Leid, Mark; Moore, Frank L.; Murray, Thomas F.

In: Journal of Neurochemistry, Vol. 75, No. 1, 2000, p. 413-423.

Research output: Contribution to journalArticle

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