At normal conditions there is a substantial water gradient over the skin as it separates the water-rich inside of the body from the dry outside. This leads to a variation in the degree of hydration along the skin and changes in this gradient may affect the structure and function of skin. In this study we raise the question: How do changes in the water gradient across skin affect its permeability? We approach this problem in experiments that permit strict control of the gradient in the chemical potential of water. The results demonstrate that an external water gradient can be used to regulate transport of drugs across the skin. It is shown that the permeability of the skin barrier increases abruptly at low water gradients, corresponding to high degrees of skin hydration, and that this effect is reversible. This phenomenon is highly relevant to drug delivery applications due to its potential of temporarily opening the skin barrier for transdermal delivery of drugs and subsequently closing the barrier after treatment. The results are explained on basis that the skin is a responding membrane, for which small changes in the environment can lead to major changes in membrane structure, which in turn affect its transport properties. We have in parallel theoretical modeling and experimental studies in model systems shown how a water gradient across multilayer lipid membrane can be used as a regulating mechanism to control the barrier properties. These principles are here applied to the barrier of stratum corneum, the upper layer of the human skin, where it can provide an explanation for the experimental findings that a water gradient can be used to regulate drug transport across the skin.