Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes

Emma E. Johnson, Helen Gibson, Beverley Nicol, Johannes Zanzinger, Peter Widdowson, Mark Hawthorn, Géza Toth, J. Farkas, Remo Guerrini, David G. Lambert

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Nociceptin/orphanin FQ (N/OFQ) is the endogenous ligand for the N/OFQ receptor (NOP), whose characteristics in the dog are unknown. We therefore compared [3H]N/OFQ binding in dog and rat brain membranes. Radioligand saturation/competition studies with these membranes and leucyl-[3H]N/OFQ(1-17)OH or the novel radioligand [ 3H]N/OFQ(1-13)NH2 were performed to determine receptor density and ligand affinity. The density of classic opioid receptors was determined by using [3H]diprenorphine. Leucyl-[ 3H]N/OFQ(1-17)OH binding was concentration dependent and saturable in dog (maximum binding capacity [Bmax], 28.7 ± 2.8 fmol/mg of protein; equilibrium dissociation constant as negative log [pKd], 10.27 ± 0.11) and rat (Bmax, 137.0 ± 12.9 fmol/mg of protein; pKd, 10.41 ± 0.05). In comparison, the B max and pKd of [3H]diprenorphine were, respectively, 77.7 ± 5.3 fmol/mg of protein and 9.74 ± 0.09 in dog and 79.1 ± 18.2 fmol/mg of protein and 9.51 ± 0.04 in rat. In dog, [3H]N/OFQ(1-13)NH2 binding to NOP receptors was also saturable (Bmax, 23.7 ± 2.0 fmol/mg of protein; pK d, 10.16 ± 0.12). In both species, leucyl-[3H]N/ OFQ(1-17)OH was displaced by various NOP ligands. Dynorphin A, N/OFQ(1-5)NH 2, and nocistatin were essentially inactive. There was a significant positive correlation (r2 = 0.95; P <0.0001) between pK i values (an estimate of affinity) obtained in displacement studies in rat and dog. We have demonstrated a low density of NOP receptors, measured with two radioligands, in dog, and these receptors display a high degree of pharmacological similarity with those natively expressed in the rat.

Original languageEnglish
Pages (from-to)741-747
Number of pages7
JournalAnesthesia and Analgesia
Volume97
Issue number3
Publication statusPublished - Sep 1 2003

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Binding Sites
Dogs
Membranes
Brain
Diprenorphine
Ligands
Proteins
Dynorphins
Opioid Receptors
nociceptin
Pharmacology
nociceptin receptor
nociceptin orphanin FQ(1-17)OH

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

Johnson, E. E., Gibson, H., Nicol, B., Zanzinger, J., Widdowson, P., Hawthorn, M., ... Lambert, D. G. (2003). Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes. Anesthesia and Analgesia, 97(3), 741-747.

Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes. / Johnson, Emma E.; Gibson, Helen; Nicol, Beverley; Zanzinger, Johannes; Widdowson, Peter; Hawthorn, Mark; Toth, Géza; Farkas, J.; Guerrini, Remo; Lambert, David G.

In: Anesthesia and Analgesia, Vol. 97, No. 3, 01.09.2003, p. 741-747.

Research output: Contribution to journalArticle

Johnson, EE, Gibson, H, Nicol, B, Zanzinger, J, Widdowson, P, Hawthorn, M, Toth, G, Farkas, J, Guerrini, R & Lambert, DG 2003, 'Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes', Anesthesia and Analgesia, vol. 97, no. 3, pp. 741-747.
Johnson EE, Gibson H, Nicol B, Zanzinger J, Widdowson P, Hawthorn M et al. Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes. Anesthesia and Analgesia. 2003 Sep 1;97(3):741-747.
Johnson, Emma E. ; Gibson, Helen ; Nicol, Beverley ; Zanzinger, Johannes ; Widdowson, Peter ; Hawthorn, Mark ; Toth, Géza ; Farkas, J. ; Guerrini, Remo ; Lambert, David G. / Characterization of nociceptin/orphanin FQ binding sites in dog brain membranes. In: Anesthesia and Analgesia. 2003 ; Vol. 97, No. 3. pp. 741-747.
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