Aquaporin 5 (AQP5) implicated in the generation of saliva, tears, and pulmonary secretions functions as a water-specific channel. Epididymal epithelial cells actively reabsorb water, ions and proteins. Large quantity of testicular fluid movement across the epididymal tubules generates high osmotic milieu which is important for sperm maturation. In an effort to understand the fluid homeostasis and its regulation by sex steroids in male reproductive tract, the expression of AQP5 was examined in different regions of mouse epididymis during postnatal development. The effect of androgen on the expression of epididymal AQP5 was examined in ORX model. AQP5 mRNA levels were the highest in corpus region in which drastic increase was noted during sexual maturation. Epididymal AQP5 immunoreactivity was largely found in apical as well as basal region of luminal epithelia. Moderate immunoreactivity for AQP5 was found in in smooth muscle cells in both immature and mature mice. Epididymal lumen of ORX mice showed shrinkage together with decrease in AQP5 expression. Alteration of AQP5 expression in ORX epididymis was partially recovered by androgen injection. AQP5 mRNA was induced at 10uM 5α-DHT in organ cultured epididymis. Chromatin immunoprecipitatin (ChIP) showed that 5α-DHT induced recruitment of androgen receptor (AR) to the -4635 to -4453 bp region of the AQP5 gene promoter in adult epididymis. Taken together, axial regulatory mechanism may control transcription of AQP5 along the length of epididymal tubule. Water transport through AQP5 is important for late sperm maturation and storage in epididymis. Androgen may directly induce AQP5 gene transcription via activation of AR in epididymis.