This study investigated the response of isolated tracheal preparation (ITP) from guinea pigs to three bronchoconstrictive drugs: histamine (H), carbachol (CAR), and acetylcholine (ACH). Cumulative dose-response curves were obtained in substrate-rich (5.50 mM of glucose) and substrate-free physiological salt solutions under aerobic conditions (PO2 = 95 +/- 0.5 Torr) or in acute 30-min hypoxia (PO2 = 30 +/- 2 Torr). The reactivity of ITP was measured by ED50 (effective dose developing 50% of maximal response); and the contractility of ITP was measured by a maximal developed isometric tension. This study showed that the contractile response of isolated segments of tracheas to H, CAR, and ACH was significantly decreased (P less than 0.05) when the experimental medium contained no glucose, or when glucose was replaced with metabolic inhibitor 2-deoxyglucose. This data contrasted with findings obtained with main pulmonary arteries (MPAs) isolated from the same species, where the absence of glucose in the experimental medium did not affect patterns of histamine cumulative dose-response curves. It was also shown that pretreatment of isolated tracheas with a small dose of insulin significantly decreased (P less than 0.05) the contractile response of airway smooth muscle to histamine. In both experiments, either with or without glucose in the experimental medium, acute hypoxia (30 min) had a significantly greater effect (P less than 0.05) on the development of isometric tension of MPA compared with that on ITP. Both removal of glucose from the experimental medium and/or acute hypoxia significantly decreased (P less than 0.001) adenosin triphosphate and creatine phosphate contents of tracheal segments but did not influence the high-energy phosphate content of main pulmonary arteries exposed to similar conditions. In addition to pointing out the physiological and metabolic differences between pulmonary vascular and airway smooth muscle, this study showed the important dependence of airway smooth muscle on glucose as a substrate.