Maternal care of pups influences the development of hypothalamic-pituitary-adrenal (HPA) responses to stress in the rat. Thus, the adult offspring of mothers that naturally show an increased frequency of pup licking/grooming and arched back nursing (ie, High LG-ABN mothers) show more modest HPA responses to stress. These effects are, in part, mediated by changes in hippocampal glucocorticoid receptor (GR) gene expression. We propose a working model describing the cellular and molecular basis for this maternal effect. The changes in maternal care increase serotonin (5-HT) turnover in the hippocampus which, in turn, activates cAMP formation via a 5-HT7 receptor and to increase protein kinase A activity and increased expression of activator protein-2 (AP-2) and NGFI-A which transactivate GR gene transcription via direct interaction with relevant GR gene promoter sequences. The critical questions concerns the apparent perminance of these effects: How might maternal care program differences in gene expression over the lifespan of the offspring. Our studies are designed to examine hypothesis that such effects are mediated by structural changes in DNA sequences located at specific sites within the promoter region of the GR gene. These studies use both in vivo and in vitro approaches to examine how maternal care alters DNA methylation and the relationship between such chemical alterations of the DNA, chromatin structure, gene expression and physiology. The methods include analysis of single nucleotide methylation using sodium bisulfite mapping, histone modifications and DNA-protein interactions with chromatin immunoprecipitation assays. We believe that these studies hold the possibility of providing a clear description of the mechanisms by which maternal care exerts long-term effects over gene expression in the brain and thus of phenotypic development. In essence, these studies directly examine gene X environment interactions in relation to specific, functional trait. Finally, although normally loath to use such descriptions, to the best of our knowledge these studies potentially provide the first description of structural changes in DNA in response to a post-mitotic, environmental event, and may ultimately provide the initial studies of environmentally-induced plasticity in DNA methyation and chromatin structure over the lifespan.