Previous studies indicate an important role for estrogen in memory and learning. Aromatase-knockout (ArKO) mice are unable to produce estrogen because they lack a functional Cyp 19 gene that encodes for aromatase, the enzyme that converts testosterone into estrogen. Using a Y-maze test for short-term spatial reference memory, we found that both male and female ArKO mice performed significantly worse than wildtype controls. Gonadectomy reduced Y-maze responses in male and female wildtype controls, but had no effect in ArKO mice. After gonadectomy, there was no significant difference between wildtype and ArKO mice. For the first time using ArKO mice, our findings confirm the importance of estrogen in memory in both males and females.