It is predicted that between 2015 to 2050, that the world aged population will nearly double from 12% to 22%. Associated with this increase will be high risk of experiencing negative effects of ageing, including common diseases occurring with advanced age such as Alzheimer’s disease and cancer. Epigenetic alterations across the course of ageing are known to have a strong association with these chronic diseases. However, a high-quality condition for living, called environmental enrichment (EE), can rescue epigenetic changes that are linked to age-related neuronal impairment, resulting in the reduction of pathological risk during ageing. Most studies of epigenetic changes in the ageing brain use whole brain homogenate, and three studies have examined purified neurons. This is a limitation because different cell and neuron subtypes have different epigenetic signatures. The aim of this study is to generate the first genome-wide data investigating epigenetic signatures in excitatory neurons from aged mice exposed to EE versus standard housing conditions. To achieve this aim, male C57/BL6 wild-type 12, 18- and 24-month-old mice were aged in standard housing and EE conditions (n=10 per age/housing condition). At 11, 17 and 23 months of age the cohorts of mice were cognitively tested (open field, Y maze, Barnes maze). A novel FACS protocol was used to purify excitatory neurons, then NOMe-seq was used to explore DNA methylation and nucleosome positioning changes across ageing with standard housing or EE. Differentially methylated regions and nucleosome depleted regions altered across the aging time course in excitatory neurons (p<0.05). Additionally, lifelong exposure to an enriched environment significantly improved the DNA methylation and nucleosome landscape with age (p<0.05). The comprehensive picture of age-environment interactions in animal models may aid in developing potential anti-cancer strategies and the reinstatement of learning and memory function in neurodegenerative conditions.