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Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells

BACKGROUND: Alcoholism is a widespread chronic disorder of complex aetiology with a significant negative impact on the individual and the society. Mechanisms of ethanol action are not sufficiently well understood at the molecular level and the pharmacotherapy of alcoholism is still in its infancy. O...

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Autores principales: Vukojević, Vladana, Ming, Yu, D'Addario, Claudio, Rigler, Rudolf, Johansson, Björn, Terenius, Lars
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2602977/
https://www.ncbi.nlm.nih.gov/pubmed/19104662
http://dx.doi.org/10.1371/journal.pone.0004008
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author Vukojević, Vladana
Ming, Yu
D'Addario, Claudio
Rigler, Rudolf
Johansson, Björn
Terenius, Lars
author_facet Vukojević, Vladana
Ming, Yu
D'Addario, Claudio
Rigler, Rudolf
Johansson, Björn
Terenius, Lars
author_sort Vukojević, Vladana
collection PubMed
description BACKGROUND: Alcoholism is a widespread chronic disorder of complex aetiology with a significant negative impact on the individual and the society. Mechanisms of ethanol action are not sufficiently well understood at the molecular level and the pharmacotherapy of alcoholism is still in its infancy. Our study focuses at the cellular and molecular level on ethanol-induced effects that are mediated through the mu-opioid receptor (MOP) and on the effects of naltrexone, a well-known antagonist at MOP that is used clinically to prevent relapse in alcoholism. METHODOLOGY/PRINCIPAL FINDINGS: Advanced fluorescence imaging by Confocal Laser Scanning Microscopy (CLSM) and Fluorescence Correlation Spectroscopy (FCS) are used to study ethanol effects on MOP and plasma membrane lipid dynamics in live PC12 cells. We observed that relevant concentrations of ethanol (10–40 mM) alter MOP mobility and surface density, and affect the dynamics of plasma membrane lipids. Compared to the action of specific ligands at MOP, ethanol-induced effects show complex kinetics and point to a biphasic underlying mechanism. Pretreatment with naloxone or naltrexone considerably mitigates the effects of ethanol. CONCLUSIONS/SIGNIFICANCE: We suggest that ethanol acts by affecting the sorting of MOP at the plasma membrane of PC12 cells. Naltrexone exerts opposite effects on MOP sorting at the plasma membrane, thereby countering the effects of ethanol. Our experimental findings give new insight on MOP-mediated ethanol action at the cellular and molecular level. We suggest a new hypothesis to explain the well established ethanol-induced increase in the activity of the endogenous opioid system.
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spelling pubmed-26029772008-12-23 Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells Vukojević, Vladana Ming, Yu D'Addario, Claudio Rigler, Rudolf Johansson, Björn Terenius, Lars PLoS One Research Article BACKGROUND: Alcoholism is a widespread chronic disorder of complex aetiology with a significant negative impact on the individual and the society. Mechanisms of ethanol action are not sufficiently well understood at the molecular level and the pharmacotherapy of alcoholism is still in its infancy. Our study focuses at the cellular and molecular level on ethanol-induced effects that are mediated through the mu-opioid receptor (MOP) and on the effects of naltrexone, a well-known antagonist at MOP that is used clinically to prevent relapse in alcoholism. METHODOLOGY/PRINCIPAL FINDINGS: Advanced fluorescence imaging by Confocal Laser Scanning Microscopy (CLSM) and Fluorescence Correlation Spectroscopy (FCS) are used to study ethanol effects on MOP and plasma membrane lipid dynamics in live PC12 cells. We observed that relevant concentrations of ethanol (10–40 mM) alter MOP mobility and surface density, and affect the dynamics of plasma membrane lipids. Compared to the action of specific ligands at MOP, ethanol-induced effects show complex kinetics and point to a biphasic underlying mechanism. Pretreatment with naloxone or naltrexone considerably mitigates the effects of ethanol. CONCLUSIONS/SIGNIFICANCE: We suggest that ethanol acts by affecting the sorting of MOP at the plasma membrane of PC12 cells. Naltrexone exerts opposite effects on MOP sorting at the plasma membrane, thereby countering the effects of ethanol. Our experimental findings give new insight on MOP-mediated ethanol action at the cellular and molecular level. We suggest a new hypothesis to explain the well established ethanol-induced increase in the activity of the endogenous opioid system. Public Library of Science 2008-12-23 /pmc/articles/PMC2602977/ /pubmed/19104662 http://dx.doi.org/10.1371/journal.pone.0004008 Text en Vukojević et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Vukojević, Vladana
Ming, Yu
D'Addario, Claudio
Rigler, Rudolf
Johansson, Björn
Terenius, Lars
Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title_full Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title_fullStr Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title_full_unstemmed Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title_short Ethanol/Naltrexone Interactions at the mu-Opioid Receptor. CLSM/FCS Study in Live Cells
title_sort ethanol/naltrexone interactions at the mu-opioid receptor. clsm/fcs study in live cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2602977/
https://www.ncbi.nlm.nih.gov/pubmed/19104662
http://dx.doi.org/10.1371/journal.pone.0004008
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