Autoradiography in opioid triple knockout mice reveals opioid and opioid receptor like binding of naloxone benzoylhydrazone

Veronica Cox, Siân Clarke, Traci Czyzyk-Morgan, Micheal Ansonoff, Joshua Nitsche, Ming Sing Hsu, Anna Borsodi, Csaba Tömböly, Géza Tóth, Ray Hill, John Pintar, Ian Kitchen

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Naloxone benzoylhydrazone (NalBzoH) is a ligand used to study opioid receptors. It has been suggested to act at a novel kappa3 receptor but also appears to bind to classical opioid receptors, and possibly the ORL1 receptor. We have used opioid receptor triple knockout mice, deficient in genes coding for the μ, δ and κ-receptor, to characterise the relative contributions of opioid and ORL1 activity to the binding of this ligand, by carrying out receptor autoradiography with [3H]NalBzoH. As competing ligands we have used diprenorphine and nociceptin at 1 μM, alone or in combination, to determine the contribution of opioid and ORL1 receptor binding. At 4 nM [3H]NalBzoH showed labelling in wild-type brains indicative of broad spectrum classical opioid receptor binding. In the triple knockout brains all labelling was completely absent, suggesting that at this concentration there is no binding to ORL1 sites. However at 50 nM [ 3H]NalBzoH showed labelling in triple knockout brains with a distribution pattern indicative of ORL1 labelling. Quantitative analysis showed that nociceptin displaced typically 30% of the residual labelling in knockout brains whilst diprenorphine had relatively little effect. The data show that at 50 nM NalBzoH no binding was detected other than to classical opioid receptors or to ORL1 in an approximate ratio of 2:1.

Original languageEnglish (US)
Pages (from-to)228-235
Number of pages8
JournalNeuropharmacology
Volume48
Issue number2
DOIs
StatePublished - Jan 1 2005

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Opioid Receptors
Autoradiography
Knockout Mice
Opioid Analgesics
Diprenorphine
Brain
Ligands
naloxone benzoylhydrazone
Genes

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Cellular and Molecular Neuroscience

Cite this

Cox, Veronica ; Clarke, Siân ; Czyzyk-Morgan, Traci ; Ansonoff, Micheal ; Nitsche, Joshua ; Hsu, Ming Sing ; Borsodi, Anna ; Tömböly, Csaba ; Tóth, Géza ; Hill, Ray ; Pintar, John ; Kitchen, Ian. / Autoradiography in opioid triple knockout mice reveals opioid and opioid receptor like binding of naloxone benzoylhydrazone. In: Neuropharmacology. 2005 ; Vol. 48, No. 2. pp. 228-235.
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abstract = "Naloxone benzoylhydrazone (NalBzoH) is a ligand used to study opioid receptors. It has been suggested to act at a novel kappa3 receptor but also appears to bind to classical opioid receptors, and possibly the ORL1 receptor. We have used opioid receptor triple knockout mice, deficient in genes coding for the μ, δ and κ-receptor, to characterise the relative contributions of opioid and ORL1 activity to the binding of this ligand, by carrying out receptor autoradiography with [3H]NalBzoH. As competing ligands we have used diprenorphine and nociceptin at 1 μM, alone or in combination, to determine the contribution of opioid and ORL1 receptor binding. At 4 nM [3H]NalBzoH showed labelling in wild-type brains indicative of broad spectrum classical opioid receptor binding. In the triple knockout brains all labelling was completely absent, suggesting that at this concentration there is no binding to ORL1 sites. However at 50 nM [ 3H]NalBzoH showed labelling in triple knockout brains with a distribution pattern indicative of ORL1 labelling. Quantitative analysis showed that nociceptin displaced typically 30{\%} of the residual labelling in knockout brains whilst diprenorphine had relatively little effect. The data show that at 50 nM NalBzoH no binding was detected other than to classical opioid receptors or to ORL1 in an approximate ratio of 2:1.",
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Cox, V, Clarke, S, Czyzyk-Morgan, T, Ansonoff, M, Nitsche, J, Hsu, MS, Borsodi, A, Tömböly, C, Tóth, G, Hill, R, Pintar, J & Kitchen, I 2005, 'Autoradiography in opioid triple knockout mice reveals opioid and opioid receptor like binding of naloxone benzoylhydrazone', Neuropharmacology, vol. 48, no. 2, pp. 228-235. https://doi.org/10.1016/j.neuropharm.2004.09.016

Autoradiography in opioid triple knockout mice reveals opioid and opioid receptor like binding of naloxone benzoylhydrazone. / Cox, Veronica; Clarke, Siân; Czyzyk-Morgan, Traci; Ansonoff, Micheal; Nitsche, Joshua; Hsu, Ming Sing; Borsodi, Anna; Tömböly, Csaba; Tóth, Géza; Hill, Ray; Pintar, John; Kitchen, Ian.

In: Neuropharmacology, Vol. 48, No. 2, 01.01.2005, p. 228-235.

Research output: Contribution to journalArticle

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