Effects of trichostatin A on neuronal mu-opioid receptor gene expression

Ying Chih Lin, Kelly E. Flock, Ryan J. Cook, Amanda J. Hunkele, Horace H. Loh, Jane L. Ko

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

In this study, we determined the effects of a histone deacetylase (HDAC) inhibitor, trichostatin A (TSA), on neuronal mu-opioid receptor (MOR) gene expression using human neuronal NMB cells, endogenously expressing MOR. Recruitment of two classes of HDAC, HDAC1 and HDAC2, to MOR promoter region in situ was detected via chromatin immunoprecipitation (ChIP) analysis with NMB cells. Functional analysis using the luciferase reporter gene system showed that TSA induced an approximately 3-fold increase of the promoter activity as compared to the vehicle treated group. Mutation analysis demonstrated that TSA response was mediated by both dsDNA (Sp1/Sp3 binding site) and ssDNA (PolyC binding protein1, PCBP, binding site) elements located in mouse MOR proximal core promoter region, further suggesting the functional importance of this cis-element, which shows high sequence homology between human and mouse MOR genes. ChIP analysis further suggested that TSA enhanced the recruitment of Sp1/Sp3 and PCBP to the promoter region, whereas no significant changes of total proteins were observed in response to TSA using Western blot analysis. Moreover, confocal images showed TSA-induced nuclear hot spots of endogenous PCBP in neuronal cells, whereas no obvious nuclear PCBP hotspot was observed in vehicle treated cells. Taken together, these results suggested that TSA enhanced neuronal MOR gene expression at the transcriptional level. RT-PCR analysis further revealed that TSA also decreased the steady-state level of MOR mRNA in a time-dependent manner by enhancing its instability. Thus, data suggest that TSA, an epigenetic regulator, affects neuronal MOR gene expression at both transcriptional and post-transcriptional levels.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalBrain Research
Volume1246
DOIs
StatePublished - Dec 30 2008

Bibliographical note

Funding Information:
The acquisition of Fluoview 1000 confocal microscope was made possible by a MRI grant of NSF. I thank Drs. Hsien-Ching Liu and Andrew P. Smith for editing the manuscript. This research was supported by NIH research grant DA-016673.

Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.

Keywords

  • Neuronal MOR gene expression
  • Proximal promoter
  • Sp1/Sp3 and PCBP factors
  • Transcriptional activation
  • Trichostatin A
  • mRNA degradation

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