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Chemotherapy‐associated memory deficits in adults are prevalent with systemic treatment utilizing 5‐fluorouracil (5‐Fu). 5‐Fu disrupts cell proliferation and readily crosses the blood–brain barrier. Proliferating cells within the adult dentate gyrus of the hippocampus give rise to new neurons involved in memory and learning and require neurotrophic factors such as brain‐derived neurotrophic factor (BDNF) to nurture this process of adult neurogenesis. Some of these proliferating cells are anatomically and functionally supported by vascular endothelial cells. We propose that systemically administered 5‐Fu chemotherapy will cause deficits in hippocampal memory that are associated with altered BDNF levels and proliferating cells (particularly vascular‐associated cells) in the dentate gyrus. This was tested by determining the effect of 5‐Fu on spatial working memory as modelled by the object location recognition test …