In vitro studies have contributed to the characterization of receptors in cranial blood vessels and the identification of possible new antimigraine agents. Animal models enable the study of vascular responses, neurogenic inflammation, and peptide release, and thus have provided leads in the search for migraine mechanisms. So far, however, animal models cannot predict the efficacy of new therapies for migraine. Because migraine attacks are fully reversible and can be aborted by therapy, the headache- or migraine-provoking property of naturally occurring signaling molecules can be tested in a human model. If a naturally occurring substance can provoke migraine in human patients, then it is likely, although not certain, that blocking its effect will be effective in the treatment of acute migraine attacks. To this end, a human in vivo model of experimental headache and migraine in humans has been developed. This model has predicted the efficacy of nitric oxide synthase inhibition and calcitonin gene-related peptide receptor blockade, and has been used to examine other endogenous signaling molecules as well as genetic susceptibility factors.