Epigenetic regulation of estrogen receptor α gene expression in the mouse cortex during early postnatal development

Estrogens play a critical role in brain development by acting on areas that express estrogen receptors. In the rodent cortex, estrogen receptor α (ERα) mRNA expression is high early in postnatal development but declines starting at postnatal day (PND) 10 and is virtually absent in the adult cortex. The mechanisms controlling this regulation are largely unknown. Methylation is important for gene silencing during development in many tissues, including the brain. In the present study, we examined the methylation status of ERα 5′ untranslated exons during early postnatal development in male and female mice using methylation-specific PCR and pyrosequencing. Several regions of ERα promoter displayed a significant increase in methylation at PND 18 and 25 compared with PND 4. DNA methyltransferases (DNMT) are important for the initiation and maintenance of methylation. Real-time PCR showed that DNMT3A, the de novo DNMT peaked at PND 10 and was decreased by PND 25. DNMT1, which is important for maintenance of methylation, increased across development and stayed high in adult cortex. The methyl-CpG-binding protein 2 (MeCP2) is also important for stabilization of methylation. A chromatin immunoprecipitation assay showed a correlation between association of MeCP2 with ERα promoter and the increase in methylation and decrease in ERαexpression after PND 10. In mice containing a mutant MeCP2 protein, ERαmRNA expression and promoter methylation patterns across development were different compared with wild-type mice. These data suggest that methylation of ERα promoters regulates ERαmRNA expression in the cortex during postnatal development in a MeCP2-dependent fashion.