Methylation of migraine-related genes in different tissues of the rat

PLoS One. 2014 Mar 7;9(3):e87616. doi: 10.1371/journal.pone.0087616. eCollection 2014.


17ß-Estradiol, an epigenetic modulator, is involved in the increased prevalence of migraine in women. Together with the prophylactic efficacy of valproate, which influences DNA methylation and histone modification, this points to the involvement of epigenetic mechanisms. Epigenetic studies are often performed on leukocytes, but it is unclear to what extent methylation is similar in other tissues. Therefore, we investigated methylation of migraine-related genes that might be epigenetically regulated (CGRP-ergic pathway, estrogen receptors, endothelial NOS, as well as MTHFR) in different migraine-related tissues and compared this to methylation in rat as well as human leukocytes. Further, we studied whether 17ß-estradiol has a prominent role in methylation of these genes. Female rats (n = 35) were ovariectomized or sham-operated and treated with 17β-estradiol or placebo. DNA was isolated and methylation was assessed through bisulphite treatment and mass spectrometry. Human methylation data were obtained using the Illumina 450k genome-wide methylation array in 395 female subjects from a population-based cohort study. We showed that methylation of the Crcp, Calcrl, Esr1 and Nos3 genes is tissue-specific and that methylation in leukocytes was not correlated to that in other tissues. Interestingly, the interindividual variation in methylation differed considerably between genes and tissues. Furthermore we showed that methylation in human leukocytes was similar to that in rat leukocytes in our genes of interest, suggesting that rat may be a good model to study human DNA methylation in tissues that are difficult to obtain. In none of the genes a significant effect of estradiol treatment was observed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcitonin Receptor-Like Protein / genetics
  • DNA Methylation / genetics
  • DNA Methylation / physiology
  • Epigenesis, Genetic / genetics
  • Estradiol / metabolism
  • Estrogen Receptor alpha / genetics
  • Female
  • Humans
  • Leukocytes / metabolism
  • Migraine Disorders / genetics*
  • Nitric Oxide Synthase Type III / genetics
  • Ovariectomy
  • Rats
  • Receptors, Calcitonin Gene-Related Peptide / genetics
  • Receptors, Estrogen / genetics


  • CALCRL protein, human
  • Calcitonin Receptor-Like Protein
  • ESR1 protein, human
  • Estrogen Receptor alpha
  • Receptors, Calcitonin Gene-Related Peptide
  • Receptors, Estrogen
  • Estradiol
  • Nitric Oxide Synthase Type III

Grant support

The generation and management of genomics data for the Rotterdam Study is supported by the Netherlands Organisation of Scientific Research NWO investments (nr. 175.010.2005.011, 911-03-012) and the Netherlands Genomics Initiative (NGI)/NWO project nr. 050-060-810 (Netherlands Consortium for Healthy Aging; NCHA). The Rotterdam Study is funded by the Erasmus Medical Center and Erasmus University, Rotterdam, Netherlands Organisation for the Health Research and development (ZonMw), the Research Institute for Diseases in the Elderly (RIDE), the Ministry of Education, Culture and Science, the Ministry for Health, Welfare and Sports, the European Commission (DG XII) and the Municipality of Rotterdam. Dr. Antoinette MaassenVanDenBrink was supported by the Netherlands Organization for Scientific Research (Vidi grant 917-11-349). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.