Inhibition of forebrain μ-opioid receptor signaling by low concentrations of rimonabant does not require cannabinoid receptors and directly involves μ-opioid receptors

Ferenc Zádor, F. Ötvös, S. Benyhe, Andreas Zimmer, Eszter Páldy

Research output: Article

17 Citations (Scopus)

Abstract

Increasing number of publications shows that cannabinoid receptor 1 (CB1) specific compounds might act in a CB1 independent manner, including rimonabant, a potent CB1 receptor antagonist. Opioids, cannabinoids and their receptors are well known for their overlapping pharmacological properties. We have previously reported a prominent decrease in μ-opioid receptor (MOR) activity when animals were acutely treated with the putative endocannabinoid noladin ether (NE). In this study, we clarified whether the decreased MOR activation caused by NE could be reversed by rimonabant in CB1 receptor deficient mice. In functional [35S] GTPγS binding assays, we have elucidated that 0.1 mg/kg of intraperitoneal (i.p.) rimonabant treatment prior to that of NE treatment caused further attenuation on the maximal stimulation of Tyr-d-Ala-Gly-(NMe)Phe-Gly-ol (DAMGO), which is a highly specific MOR agonist. Similar inhibitory effects were observed when rimonabant was injected i.p. alone and when it was directly applied to forebrain membranes. These findings are cannabinoid receptor independent as rimonabant caused inhibition in both CB1 single knockout and CB1/CB2 double knockout mice. In radioligand competition binding assays we highlighted that rimonabant fails to displace effectively [3H]DAMGO from MOR in low concentrations and is highly unspecific on the receptor at high concentrations in CB1 knockout forebrain and in their wild-type controls. Surprisingly, docking computational studies showed a favorable binding position of rimonabant to the inactive conformational state of MOR, indicating that rimonabant might behave as an antagonist at MOR. These findings were confirmed by radioligand competition binding assays in Chinese hamster ovary cells stably transfected with MOR, where a higher affinity binding site was measured in the displacement of the tritiated opioid receptor antagonist naloxone. However, based on our in vivo data we suggest that other, yet unidentified mechanisms are additionally involved in the observed effects.

Original languageEnglish
Pages (from-to)378-388
Number of pages11
JournalNeurochemistry International
Volume61
Issue number3
DOIs
Publication statusPublished - aug. 2012

Fingerprint

rimonabant
Cannabinoid Receptors
Opioid Receptors
Prosencephalon
Ala(2)-MePhe(4)-Gly(5)-enkephalin
Cannabinoid Receptor CB2
Endocannabinoids
Narcotic Antagonists
Naloxone
Cricetulus

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

Cite this

@article{99a6be117931485a89feea650e85e151,
title = "Inhibition of forebrain μ-opioid receptor signaling by low concentrations of rimonabant does not require cannabinoid receptors and directly involves μ-opioid receptors",
abstract = "Increasing number of publications shows that cannabinoid receptor 1 (CB1) specific compounds might act in a CB1 independent manner, including rimonabant, a potent CB1 receptor antagonist. Opioids, cannabinoids and their receptors are well known for their overlapping pharmacological properties. We have previously reported a prominent decrease in μ-opioid receptor (MOR) activity when animals were acutely treated with the putative endocannabinoid noladin ether (NE). In this study, we clarified whether the decreased MOR activation caused by NE could be reversed by rimonabant in CB1 receptor deficient mice. In functional [35S] GTPγS binding assays, we have elucidated that 0.1 mg/kg of intraperitoneal (i.p.) rimonabant treatment prior to that of NE treatment caused further attenuation on the maximal stimulation of Tyr-d-Ala-Gly-(NMe)Phe-Gly-ol (DAMGO), which is a highly specific MOR agonist. Similar inhibitory effects were observed when rimonabant was injected i.p. alone and when it was directly applied to forebrain membranes. These findings are cannabinoid receptor independent as rimonabant caused inhibition in both CB1 single knockout and CB1/CB2 double knockout mice. In radioligand competition binding assays we highlighted that rimonabant fails to displace effectively [3H]DAMGO from MOR in low concentrations and is highly unspecific on the receptor at high concentrations in CB1 knockout forebrain and in their wild-type controls. Surprisingly, docking computational studies showed a favorable binding position of rimonabant to the inactive conformational state of MOR, indicating that rimonabant might behave as an antagonist at MOR. These findings were confirmed by radioligand competition binding assays in Chinese hamster ovary cells stably transfected with MOR, where a higher affinity binding site was measured in the displacement of the tritiated opioid receptor antagonist naloxone. However, based on our in vivo data we suggest that other, yet unidentified mechanisms are additionally involved in the observed effects.",
keywords = "μ-Opioid receptor, [ S]GTPγS binding, CB cannabinoid receptor, CB receptor knockout mouse, CB/CB receptor double knockout mouse, Docking, Forebrain, Radioligand binding, Rimonabant",
author = "Ferenc Z{\'a}dor and F. {\"O}tv{\"o}s and S. Benyhe and Andreas Zimmer and Eszter P{\'a}ldy",
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T1 - Inhibition of forebrain μ-opioid receptor signaling by low concentrations of rimonabant does not require cannabinoid receptors and directly involves μ-opioid receptors

AU - Zádor, Ferenc

AU - Ötvös, F.

AU - Benyhe, S.

AU - Zimmer, Andreas

AU - Páldy, Eszter

PY - 2012/8

Y1 - 2012/8

N2 - Increasing number of publications shows that cannabinoid receptor 1 (CB1) specific compounds might act in a CB1 independent manner, including rimonabant, a potent CB1 receptor antagonist. Opioids, cannabinoids and their receptors are well known for their overlapping pharmacological properties. We have previously reported a prominent decrease in μ-opioid receptor (MOR) activity when animals were acutely treated with the putative endocannabinoid noladin ether (NE). In this study, we clarified whether the decreased MOR activation caused by NE could be reversed by rimonabant in CB1 receptor deficient mice. In functional [35S] GTPγS binding assays, we have elucidated that 0.1 mg/kg of intraperitoneal (i.p.) rimonabant treatment prior to that of NE treatment caused further attenuation on the maximal stimulation of Tyr-d-Ala-Gly-(NMe)Phe-Gly-ol (DAMGO), which is a highly specific MOR agonist. Similar inhibitory effects were observed when rimonabant was injected i.p. alone and when it was directly applied to forebrain membranes. These findings are cannabinoid receptor independent as rimonabant caused inhibition in both CB1 single knockout and CB1/CB2 double knockout mice. In radioligand competition binding assays we highlighted that rimonabant fails to displace effectively [3H]DAMGO from MOR in low concentrations and is highly unspecific on the receptor at high concentrations in CB1 knockout forebrain and in their wild-type controls. Surprisingly, docking computational studies showed a favorable binding position of rimonabant to the inactive conformational state of MOR, indicating that rimonabant might behave as an antagonist at MOR. These findings were confirmed by radioligand competition binding assays in Chinese hamster ovary cells stably transfected with MOR, where a higher affinity binding site was measured in the displacement of the tritiated opioid receptor antagonist naloxone. However, based on our in vivo data we suggest that other, yet unidentified mechanisms are additionally involved in the observed effects.

AB - Increasing number of publications shows that cannabinoid receptor 1 (CB1) specific compounds might act in a CB1 independent manner, including rimonabant, a potent CB1 receptor antagonist. Opioids, cannabinoids and their receptors are well known for their overlapping pharmacological properties. We have previously reported a prominent decrease in μ-opioid receptor (MOR) activity when animals were acutely treated with the putative endocannabinoid noladin ether (NE). In this study, we clarified whether the decreased MOR activation caused by NE could be reversed by rimonabant in CB1 receptor deficient mice. In functional [35S] GTPγS binding assays, we have elucidated that 0.1 mg/kg of intraperitoneal (i.p.) rimonabant treatment prior to that of NE treatment caused further attenuation on the maximal stimulation of Tyr-d-Ala-Gly-(NMe)Phe-Gly-ol (DAMGO), which is a highly specific MOR agonist. Similar inhibitory effects were observed when rimonabant was injected i.p. alone and when it was directly applied to forebrain membranes. These findings are cannabinoid receptor independent as rimonabant caused inhibition in both CB1 single knockout and CB1/CB2 double knockout mice. In radioligand competition binding assays we highlighted that rimonabant fails to displace effectively [3H]DAMGO from MOR in low concentrations and is highly unspecific on the receptor at high concentrations in CB1 knockout forebrain and in their wild-type controls. Surprisingly, docking computational studies showed a favorable binding position of rimonabant to the inactive conformational state of MOR, indicating that rimonabant might behave as an antagonist at MOR. These findings were confirmed by radioligand competition binding assays in Chinese hamster ovary cells stably transfected with MOR, where a higher affinity binding site was measured in the displacement of the tritiated opioid receptor antagonist naloxone. However, based on our in vivo data we suggest that other, yet unidentified mechanisms are additionally involved in the observed effects.

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KW - CB/CB receptor double knockout mouse

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KW - Radioligand binding

KW - Rimonabant

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