Estrogen receptor (ER), which mediates the multiple effects of estrogen in brain, is regulated by several factors including its own ligand. In the present study, we have examined the effect of age, sex and gonadal steroids (estrogen and testosterone) on the level of ERalpha and ERbeta in the cerebral cortex of AKR mice. Adult and old mice of both sexes were divided into four groups: intact, gonadectomized, 17beta-estradiol treated and testosterone treated. Western blot analysis showed higher level of ERalpha and ERbeta in the cerebral cortex of adult female than male mice. ERbeta level decreased significantly with advancing age in both sexes, whereas 17beta-estradiol supplementation decreased ERalpha level in old male and increased in old female, it also increased ERbeta level in old male and adult female. On the other hand, testosterone treatment decreased ERalpha level significantly in old female and ERbeta level in adult female but increased ERbeta level in male mice of both ages. Thus, these findings showed that the expression of ERalpha and ERbeta protein is differentially influenced by age, sex and gonadal steroids in the mouse cerebral cortex, suggesting differences in ER-mediated brain functions.