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. Our study focuses at the cellular and molecular level on ethanol-induced effects that are mediated through the micro-opioid receptor (MOP) and on the effects of naltrexone, a well-known antagonist at MOP that is used clinically to prevent relapse in alcoholism.


RESULTS
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
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.
Vukojević
Vladana
V
Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden. vladana.vukojevic@ki.se
Ming
Yu
Y
D'Addario
Claudio
C
Rigler
Rudolf
R
Johansson
Björn
B
Terenius
Lars
L
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D000431:Ethanol / Q000378:metabolism / Q000494:pharmacology*; D008563:Membrane Lipids / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D009271:Naltrexone / Q000378:metabolism / Q000494:pharmacology*; D009292:Narcotic Antagonists / Q000494:pharmacology; D016716:PC12 Cells; D051381:Rats; D017450:Receptors, Opioid, mu / Q000187:drug effects / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors
D008563:Membrane Lipids; D009292:Narcotic Antagonists; D017450:Receptors, Opioid, mu; D000431:Ethanol; D009271:Naltrexone