Our data also suggest that the continuous intra-NAc delivery of DNMT inhibitors represses the expression of Dnmts at the transcription level in postmitotic neurons. Although DNA methylation is generally thought to be associated with transcriptional repression of the target genes, a recent study suggested that the binding of a complex of MeCP2 and cyclic AMP response element (CRE)-binding protein (CREB) to the methylated CpG site can activate transcription (Chahrour et al.,

2008). Interestingly, the putative CRE site is adjacent to CpG site 2 of the Gdnf gene ( Figure 7A). In addition, we found that MeCP2 and CREB are colocalized in the NAc ( Figure 7B). These facts led us to speculate that the binding of the MeCP2-CREB complex to the Gdnf promoter may be a causal mechanism of the increased Gdnf expression in stressed B6

mice. To test this Roxadustat manufacturer possibility, we assessed the interactions of MeCP2 and CREB in vSTR proteins of B6 and BALB mice. IP-Western blot analysis showed that there is no apparent difference in the formation of MeCP2-CREB complexes between stressed and nonstressed mice in both strains ( Figure 7C). Next, to investigate the binding of MeCP2-CREB complexes at the Gdnf promoter, we performed re-ChIP assays using an antibody for CREB on vSTR samples that had been initially immunoprecipitated with an antibody for MeCP2. Consistent with a previous report ( Chahrour et al., 2008), CREB-MeCP2 complexes on the somatostatin promoter were enriched, whereas they were reduced on the selleck chemicals myocyte enhancer factor 2c promoter (data not shown), validating the specificity of the re-ChIP used. We found that the Gdnf promoter-containing DNA fragments of stressed B6 mice were significantly enriched in the reimmunoprecipitates of samples treated with CREB antibodies compared with those of nonstressed mice. This effect was not seen in stressed BALB mice ( Figure 7D). These results L-NAME HCl suggest that the CUMS-induced binding of MeCP2-CREB complexes to the Gdnf promoter leads to the activation of its transcription. This study used genetically distinct

inbred mouse strains to describe one of the molecular mechanisms underlying susceptibility and adaptation responses to chronic stress. The proposed mechanisms underlying stress susceptibility and adaptation are described in Figure 7E. Our results suggest that CUMS increases DNA methylation at CpG site 2, and this is associated with increased MeCP2 binding. MeCP2 associated with CpG site 2 interacts with HDAC2, which in turn decreases the level of H3 acetylation and concomitantly represses Gdnf transcription, leading to the formation of a more depression-susceptible phenotype in BALB mice. Continuous IMI treatment relieves MeCP2 occupancy and reverses HDAC2 levels, which leads to normal levels of H3 acetylation and subsequent Gdnf transcription, resulting in normal emotional behaviors.