In this study, we exposed female medaka to aqueous solutions of o,p′-DDT until tissues residues reached approximately 100 g/g. Male offspring of exposed females and unexposed males that were raised to 6 months of age differed from untreated controls in their response to subsequent 4-day static exposure to 17-β-estradiol (E2; 12 μg/l as determined by steroid EIA). Homogenized liver samples from males from DDT treatments, females from DDT treatments, control males and control females produced two vitellogenin bands on a Western blot with no qualitative differences in band size and location between the groups. To quantify the relative level of vitellogenin, 5 μg of protein from each sample was analyzed by slot blot. A statistical comparison revealed a significant difference in optical density of the slots between control males and DDT exposed males (analysis of variance, n=18, P=0.021; Tukey test, n=9, P=0.047) indicating that previous transgenerational exposure to DDT enhanced the vitellogenic response to E2 exposure in males. These results indicate that exposure to estrogenic chemicals may produce an ‘estrogen memory’ which potentiates the response to a subsequent exposure event.