Devising a novel protocol to study 5-hydroxymethylcytosine's role in the consolidation of a methamphatamine [sic] associated contextual memory

DNA methylation, the addition of a methyl group to the 5' position of DNA cytosines (5mC), is generally associated with transcriptional repression during early embryo formation ; however, in the adult brain, it is dynamically regulated and plays an important role in the formation and maintenance of memory. Very recently, it has been hypothesized that DNA hydroxymethylation, the addition of a hydroxyl group to methylated cytosines (5mC), serves as an intermediate in the DNA demethylation pathway. GIven its recent discovery, the role of DNA hydroxymethylation in memory has not yet been explored. In this study, we developed an immunofluorescent triple labeling protocol in order to begin examining the involvement of 5mC and 5hmC in neurons activated by consolidation of a contextual memory associated with methamphetamine in the brain's reward center, the nucleus accumbens.