Maternal cocaine administration causes an epigenetic modification of protein kinase Cε gene expression in fetal rat heart

Haitao Zhang, Agus Darwanto, Thomas A. Linkhart, Lawrence C. Sowers, Lubo Zhang

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Protein kinase Cε (PKCε) plays a pivotal role in cardioprotection during cardiac ischemia and reperfusion injury. Recent studies demonstrated that prenatal cocaine exposure caused a decrease in PKCε expression and increased heart susceptibility to ischemic injury in adult offspring, suggesting an in utero programming of PKCε gene expression pattern in the heart. The present investigation aimed to elucidate whether an epigenetic mechanism, DNA methylation, accounts for cocaine-mediated repression of the PKCε gene in the heart. Pregnant rats were administered either saline or cocaine intraperitoneally (15 mg/kg) twice daily from days 15 to 20 of gestational age, and term fetal hearts were studied. Cocaine treatment significantly decreased PKCε mRNA and protein levels in the heart. CpG dinucleotides found in cAMP response element-binding protein (CREB), CREB/c-Jun1, and CREB/c-Jun2 binding sites at the proximal promoter region of the PKCε gene were densely methylated and were not affected by cocaine. In contrast, methylation of CpGs in the activator protein 1 (AP-1) binding sites was low but was significantly increased by cocaine. Reporter gene assays showed that the AP-1 binding site played a strong stimulatory role of PKCε gene transcription. Methylation of the AP-1 binding sites significantly decreased AP-1 binding to the PKCε promoter. Supershift analyses implicated c-Jun homodimers binding to the AP-1 binding sites. Cocaine did not affect nuclear c-Jun levels or the binding of c-Jun to the unmethylated AP-1 binding sites. The results indicate a role for DNA methylation in cocaine-mediated PKCε gene repression in the developing heart and suggest an epigenetic mechanism affecting this gene linked with vulnerability of ischemic injury in the heart of adult offspring.

Original languageEnglish (US)
Pages (from-to)1319-1328
Number of pages10
JournalMolecular pharmacology
Volume71
Issue number5
DOIs
StatePublished - May 1 2007
Externally publishedYes

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ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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