Nitric oxide and histone deacetylases modulate cocaine-induced mu-opioid receptor levels in PC12 cells.
BACKGROUND
Cocaine exposure has been reported to alter central μ-opioid receptor (MOR) expression in vivo. The present study employed an in vitro cellular model to explore possible mechanisms that may be involved in this action of cocaine.


METHODS
To assess the effects of cocaine on MOR levels, two treatment regimens were tested in PC12 cells: single continuous or multiple intermittent. MOR protein levels were assessed by western blot analysis and quantitative PCR was used to determine relative MOR mRNA expression levels. To evaluate the role of nitric oxide (NO) and histone acetylation in cocaine-induced MOR expression, cells were pre-treated with the NO synthase inhibitor Nω-nitro-L-arginine methylester (L-NAME) or the non-selective histone acetyltransferase inhibitor curcumin.


RESULTS
Both cocaine treatment regimens significantly increased MOR protein levels and protein stability, but only multiple intermittent treatments increased MOR mRNA levels as well as c-fos mRNA levels and activator protein 1 binding activity. Both regimens increased NO production, and pre-treatment with L-NAME prevented cocaine-induced increases in MOR protein and mRNA levels. Single and multiple cocaine treatment regimens inhibited histone deacetylase activity, and pre-treatment with curcumin prevented cocaine-induced up-regulation of MOR protein expression.


CONCLUSIONS
In the PC12 cell model, both NO and histone deacetylase activity regulate cocaine-induced MOR expression at both the transcriptional and post-transcriptional levels. Based on these novel findings, it is hypothesized that epigenetic mechanisms are implicated in cocaine's action on MOR expression in neurons.
Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Warren
W
Winick-Ng
Francesco
F
Leri
Bettina E
BE
Kalisch
D000818:Animals; D003042:Cocaine / Q000008:administration & dosage*; D003513:Cycloheximide / Q000494:pharmacology; D018765:Dopamine Uptake Inhibitors / Q000008:administration & dosage; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D016716:PC12 Cells; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017450:Receptors, Opioid, mu / Q000378:metabolism*
D018765:Dopamine Uptake Inhibitors; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D017450:Receptors, Opioid, mu; D009569:Nitric Oxide; D003513:Cycloheximide; D006655:Histone Deacetylases; D003042:Cocaine