Sweat secretion plays an important role in the human body. It is known that the hyperhidrosis and anhidrosis are caused by abnormalities in sweat secretion and can result in severe skin conditions such as pruritus and erythema. However, no effective therapies or therapeutic agents have yet been discovered. Previously, it is reported that Pituitary adenylate cyclase-activating polypeptide (PACAP) promotes sweating, but details of the mechanism of sweat secretion has not been clarified (Sasaki et al., 2017). Therefore, we used immortalized human eccrine gland cells to investigate how sweat secretion is induced by PACAP treatment. Intracellular Ca²⁺ levels were increased in these cells with physiological concentrations of PACAP. However, they were not elevated when cells were concomitantly treated with PA-8, PAC1-R selected antagonist with PACAP. Furthermore, results of immunocytochemistry experiments showed that aquaporin-5(AQP-5) was translocated from the cytoplasm to the cell membrane by PACAP. These results suggest that PACAP promotes sweat secretion by translocation of AQP-5 to the cell membrane in response to increased levels of intracellular Ca²⁺ on the sweat glands. These findings also provide a solid basis for future research initiatives to develop new therapies and therapeutic agents to treat sweating disorders.