AbstractThe neuropeptide galanin has been ascribed different roles in modulating physiological functions in the skin. The present study examined the function of galanin in eccrine sweat gland physiology. We demonstrated secretion of galanin by sweat glands in vivo by radioimmunoassay of human sweat (20–192 fmol galanin/ml). Furthermore, human sweat glands expressed galanin receptors GalR2 and GalR3. Using chamber short‐circuit current (Isc) measurements showed that application of galanin to human NCL‐SG3 cells led to a significant increase in Isc, which was inhibited by the presence of chloride channel blockers and in chloride‐free Krebs solution. Additionally, application of SNAP 37889, a non‐peptidergic selective antagonist of GalR3, abolished the effect of galanin on Isc. In summary, our results show that galanin can regulate transepithelial chloride ion transport and fluid secretion by stimulating GalR3 in NCL‐SG3 cells and demonstrate a possible important extraneural function of galanin in sweat gland physiology.