Studies of the responsiveness of strips of tracheal smooth muscle and the changes after sensitization of ovalbumin were carried out. The hypothesis that there might be a generalized or a selective change of airway smooth muscle responsiveness to sensitization was examined in vitro. Agonists acting on muscarinic receptors, α1-, α2-, and β-adrenoceptors, purine receptors, histamine and serotonin receptors, and leukotriene and prostaglandin receptors were tested, as well as mediators released from local nerves by field stimulation and procedures such as elevation of potassium or addition of Ca2+ ionophores which do not involve specific receptors. Sensitivity to serotonin increased significantly in sensitized animals. Total magnitude of the contraction and subsequent relaxation responses to field stimulation also increased significantly. Neither of these changes was large in magnitude. Although there were a few minor changes in sensitivity (pD2) or in maximum responses, the hypothesis of important changes in responses of any sort in tracheal muscle after sensitization was rejected. The question was raised whether this general absence of changed responsiveness in vitro reflected (i) the failure of sensitization to induce generalized smooth muscle hyperresponsiveness, (ii) the loss of the mechanisms of such responsiveness in vitro, or (iii) the inadequacy of in vitro techniques to assess responsiveness present in vivo.