Ethanol-induced changes in poly (ADP ribose) polymerase and neuronal developmental gene expression

Neuropharmacology. 2016 Nov;110(Pt A):287-296. doi: 10.1016/j.neuropharm.2016.08.001. Epub 2016 Aug 4.

Abstract

Prenatal alcohol exposure has profound effects on neuronal growth and development. Poly-ADP Ribose Polymerase (PARP) enzymes are perhaps unique in the field of epigenetics in that they directly participate in histone modifications, transcription factor modifications, DNA methylation/demethylation and are highly inducible by ethanol. It was our hypothesis that ethanol would induce PARP enzymatic activity leading to alterations in neurodevelopmental gene expression. Mouse E18 cortical neurons were treated with ethanol, PARP inhibitors, and nuclear hormone receptor transcription factor PPARγ agonists and antagonists. Subsequently, we measured PARP activity and changes in Bdnf, OKSM (Oct4, Klf4, Sox2, c-Myc), DNA methylating/demethylating factors, and Pparγ mRNA expression, promoter 5-methylcytosine (5MC) and 5-hydroxymethylcytosine (5HMC), and PPARγ promoter binding. We found that ethanol reduced Bdnf4, 9a, and Klf4 mRNA expression, and increased c-Myc expression. These changes were reversed with a PARP inhibitor. In agreement with its role in DNA demethylation PARP inhibition increased 5MC levels at the c-Myc promoter. In addition, we found that inhibition of PARP enzymatic activity increased PPARγ promoter binding, and this corresponded to increased Bdnf and Klf4 mRNA expression. Our results suggest that PARP participates in DNA demethylation and reduces PPARγ promoter binding. The current study underscores the importance of PARP in ethanol-induced changes to neurodevelopmental gene expression.

Keywords: Alcoholism; CpG; Epigenetic; Histone.

MeSH terms

  • Animals
  • Brain-Derived Neurotrophic Factor / metabolism
  • Cells, Cultured
  • DNA Methylation / drug effects
  • DNA Methylation / physiology
  • Ethanol / toxicity*
  • Fetal Alcohol Spectrum Disorders / enzymology
  • Gene Expression Regulation, Developmental / drug effects
  • Genes, Developmental / drug effects
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / metabolism
  • Mice
  • Neurons / drug effects*
  • Neurons / metabolism*
  • Neuroprotective Agents / pharmacology
  • PPAR gamma / metabolism
  • Poly(ADP-ribose) Polymerase Inhibitors / pharmacology
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Proto-Oncogene Proteins c-myc / metabolism
  • RNA, Messenger / metabolism

Substances

  • Brain-Derived Neurotrophic Factor
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • Myc protein, mouse
  • Neuroprotective Agents
  • PPAR gamma
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Proto-Oncogene Proteins c-myc
  • RNA, Messenger
  • Ethanol
  • Poly(ADP-ribose) Polymerases