Gender difference has been reported in kidney diseases including acute kidney injury (AKI). We have demonstrated that the male hormone acts as a major factor in the induction of gender difference on AKI. However, its molecular mechanisms remain to be defined. In this study, we investigated the expression of sexual hormone receptors in the mouse kidneys suffered from AKI. C57BL6 mice were subjected to either bilateral renal ischemia followed by reperfusion (I/R) or sham-operation. Androgen receptor (AR), estrogen receptor α (ERα), and estrogen receptor β (ERβ) were determined by western blot and immunohistochemical staining in the kidneys. As previously reported, I/R induced more severe renal functional and morphological damage in male mice than female mice. After sham operation, AR, ERα and ERβ were more abundant in the distal tubules and collecting ducts than the proximal tubules in both male and female mouse kidney. AR and ERα expression was greater in the male than female mice. After I/R, AR and ERα expressions significantly decreased in both male mice and female mice, with a greater decrease in the male mice than female mice. In contrast to AR and ERα, ERβ expression was not significantly different between male mice and female mice. In sham operated mouse kidneys, AR was detected in both nucleus and cytoplasm, however, in I/R-injured kidneys, AR was more strongly expressed in the nucleus than cytoplasm. ERα and ERβ expressions were shown in the same patterns with AR expression. These results indicate that the levels of AR and ERs expression are different between male and female, and their intracellular localizations were altered by I/R injury, suggesting that gender difference in I/R-induced AKI may be associated with these receptors signaling pathways.