[3H]dynorphin1-8 binding sites in frog (Rana esculenta) brain membranes

S. Benyhe, J. Simon, A. Borsodi, M. Wollemann, E. A. Barnard

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Opioid binding sites specific for [3H]dynorphin1-8 were characterized in the particulate membrane fraction of frog (Rana esculenta) brain. The degradation of the radioligand during the assay was prevented by the use of a broad spectrum of peptidase inhibitors. The binding of [3H]dynorphin1-8 to frog brain membranes was stereoselective, reversible, saturable, and displaceable by a series of opioid ligands including dynorphin1-13, bremazocine, levorphanol and naloxone. The specific binding of [3H]dynorphin1-8 can be significantly inhibited by Na+ ions and/or guanine nucleotides confirming the agonist property of the ligand in vitro. A single set of high affinity opioid binding sites with a Kd ≈ 7.5 nM is present in the membranes. The maximum density of binding sites (Bmax ≈ 1.1 pmol [3H]dynorphin1-8 per mg protein) was considerably higher than such sites in guinea-pig brain. In addition, comparison with binding of tritiated opioid peptides selective for the μ- and σ-types of opioid receptor showed that in the frog brain most of the sites labelled by [3H]dynorphin1-8 are κ-sites and that this is a rich source of such sites.

Original languageEnglish
Pages (from-to)359-364
Number of pages6
JournalNeuropeptides
Volume26
Issue number5
DOIs
Publication statusPublished - 1994

Fingerprint

Rana esculenta
Anura
Brain
Binding Sites
Opioid Analgesics
Membranes
Levorphanol
Ligands
Radioligand Assay
Guanine Nucleotides
Opioid Peptides
Opioid Receptors
Naloxone
Protease Inhibitors
Assays
Guinea Pigs
Ions
Degradation
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Clinical Neurology
  • Neuroscience(all)
  • Cellular and Molecular Neuroscience

Cite this

[3H]dynorphin1-8 binding sites in frog (Rana esculenta) brain membranes. / Benyhe, S.; Simon, J.; Borsodi, A.; Wollemann, M.; Barnard, E. A.

In: Neuropeptides, Vol. 26, No. 5, 1994, p. 359-364.

Research output: Contribution to journalArticle

Benyhe, S. ; Simon, J. ; Borsodi, A. ; Wollemann, M. ; Barnard, E. A. / [3H]dynorphin1-8 binding sites in frog (Rana esculenta) brain membranes. In: Neuropeptides. 1994 ; Vol. 26, No. 5. pp. 359-364.
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