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DNA methylation and gene expression can be altered by early life stress (ELS) and/or ethanol consumption. The present study aimed to investigate whether DNA methylation of the Vesicular Glutamate Transporters ( Vglut ) 1-3  is related to previously observed  Vglut1-3  transcriptional differences in the ventral tegmental area (VTA), nucleus accumbens (Acb), dorsal striatum (dStr) and medial prefrontal cortex (mPFC) of adult rats exposed to ELS, modelled by maternal separation, and voluntary ethanol consumption. Targeted next-generation bisulfite sequencing was performed to identify the methylation levels on 61 5′-cytosine-phosphate-guanosine-3′ sites (CpGs) in potential regulatory regions of  Vglut1 , 53 for  Vglut2 , and 51 for  Vglut3 . In the VTA, ELS in ethanol-drinking rats was associated with  Vglut1-2  CpG-specific hypomethylation, whereas bidirectional  Vglut2  methylation differences at single CpGs were associated with ELS alone. Exposure to both ELS and ethanol, in the Acb, was associated with lower promoter and higher intronic  Vglut3  methylation; and in the dStr, with higher and lower methylation in 26% and 43% of the analyzed  Vglut1  CpGs, respectively. In the mPFC, lower  Vglut2  methylation was observed upon exposure to ELS or ethanol. The present findings suggest  Vglut1-3  CpG-specific methylation signatures of ELS and ethanol drinking, underlying previously reported  Vglut1-3  transcriptional differences in the mesocorticolimbic brain.

scientific reports

DNA methylation of Vesicular Glutamate Transporters in the mesocorticolimbic brain following early-life stress and adult ethanol exposure—an explorative study