title: Genetic influences on nicotinic α5 receptor (CHRNA5) CpG methylation and mRNA expression in brain and adipose tissue.
abstract: BACKGROUND
  The nicotinic α5 receptor subunit, encoded by CHRNA5, harbors multiple functional single nucleotide polymorphisms (SNPs) that affect mRNA expression and alter the encoded protein. These polymorphisms are most notably associated with drug-taking behaviors and cognition. We previously identified common SNPs in a distant regulatory element (DRE) that increase CHRNA5 mRNA expression in the human prefrontal cortex (PFC) and confer risk for nicotine dependence. Genome-wide epigenetic studies in PFC and adipose tissue find strong effects of the DRE SNPs on CpG methylation. However, it is unclear whether DRE SNPs influence CpG methylation en route to modulating CHRNA5 mRNA expression. It is also unclear whether these polymorphisms affect expression in other brain regions, especially those mediating drug-taking behaviors.
  
  
  RESULTS
  By measuring total and allelic CHRNA5 mRNA expression in human habenula and putamen autopsy tissues, we found that CHRNA5 DRE variants considerably increase mRNA expression by up to 3.5-fold in both brain regions. Our epigenetic analysis finds no association between CpG methylation and CHRNA5 mRNA expression in the PFC or adipose tissues.
  
  
  CONCLUSIONS
  These finding suggests the mechanisms responsible for the genetic modulation of CpG methylation and mRNA expression are independent despite the DRE SNPs being highly associated with both measures. Our findings support a strong association between the DRE SNPs and mRNA expression or CpG methylation in the brain and periphery, but the independence of the two measures leads us to conclude that environmental factors affecting CpG methylation do not appear to directly modulate gene expression.
authors.affiliation: Center for Pharmacogenomics, The Ohio State University, Columbus, OH 43210 USA ; Department of Pharmacology, The Ohio State University, 5184A Graves Hall, 333. W. 10th Ave., Columbus, OH 43210 USA.
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Ramsay
authors.affiliation: National Institute of Mental Health, Human Brain Collection Core, 10 Center Drive, Rm. 4N306, Bethesda, MD USA.
authors.forename: C Harker
authors.initials: CH
authors.lastname: Rhodes
authors.affiliation: Department of Pharmacology, The Ohio State University, 5184A Graves Hall, 333. W. 10th Ave., Columbus, OH 43210 USA.
authors.forename: Keerthi
authors.initials: K
authors.lastname: Thirtamara-Rajamani
authors.affiliation: Center for Pharmacogenomics, The Ohio State University, Columbus, OH 43210 USA ; Department of Pharmacology, The Ohio State University, 5184A Graves Hall, 333. W. 10th Ave., Columbus, OH 43210 USA.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Smith
keywords: Allelic expression; Enhancer; Epigenetics; Expression quantitative trait loci; Functional polymorphism; Methylation; Nicotine; Nicotinic receptors; Repressor; Silencer; eQTL; mRNA expression