Abstract
The agonists of μ-opioid receptor (OPRM1) induce extracellular signal-regulated kinase (ERK) phosphorylation through different pathways: morphine uses the protein kinase C (PKC)-pathway, whereas fentanyl functions in a β-arrestin2-dependent manner. In addition, the two pathways result in the different cellular location of phosphorylated ERK and the activation of different sets of transcriptional factors. In the current study, the influence of the two pathways on the expression of microRNAs (miRNAs) was investigated. After treating the primary culture of rat hippocampal neurons and the mouse hippocampi with morphine or fentanyl for 3 days, seven miRNAs regulated by one or two of the agonists were identified. One of the identified miRNAs, miR-190, was down-regulated by fentanyl but not by morphine. This down-regulation was attenuated by 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene (U0126), which blocks the phosphorylation of ERK. When fentanyl-induced but not morphine-induced ERK phosphorylation was blocked in the primary cultures from β-arrestin2(−/−) mouse, fentanyl did not decrease the expression of miR-190. However, a PKC inhibitor that blocked morphine-induced ERK phosphorylation specifically had no effect on the miR-190 down-regulation. Therefore the decrease in miR-190 expression resulted from the agonist-selective ERK phosphorylation. In addition, the expressional changes in one of the miR-190 targets, neurogenic differentiation 1 (NeuroD), correlated with those in miR-190 expression, suggesting the OPRM1 could regulate the NeuroD pathways via the control of miR-190 expression.
Footnotes
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This research was supported by the National Institutes of Health National Institute on Drug Abuse [Grants DA007339, DA016674, DA000564, DA011806, K05-DA70544, K05-DA00513].
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Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.109.060848.
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ABBREVIATIONS:
- GPCR
- G protein-coupled receptor
- ERK
- extracellular signal-regulated kinase
- PKC
- protein kinase C
- OPRM1
- μ-opioid receptor
- miRNA
- microRNA
- PCR
- polymerase chain reaction
- U0126
- 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene
- Ro-31-8425
- 3-(8-(aminomethyl)-6,7,8,9-tetrahydropyrido(1,2-a)indol-10-yl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione
- PP2
- 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyramidine
- U73122
- 1-[6-[[17β-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione
- NeuroD
- neurogenic differentiation 1
- UTR
- untranslated region
- HEK
- human embryonic kidney
- CTOP
- Cys2-Tyr3-Orn5-Pen7-amide
- TIPPψ
- H-Tyr-Ticψ[CH2NH]-Phe-Phe-OH
- DMSO
- dimethyl sulfoxide
- PBS
- phosphate-buffered saline
- ANOVA
- analysis of variance
- %MPE
- percentage of maximum possible effect.
- Received September 8, 2009.
- Accepted October 23, 2009.
- © 2010 The American Society for Pharmacology and Experimental Therapeutics
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