Confinement of dogs to 10% oxygen for 14 days caused erythropoiesis and pulmonary hypertension. Histological sections of the lung tissue showed thickening of the smooth muscle component of muscular arteries and arterioles. Segments of pulmonary artery from dogs exposed to hypoxia were superfused under continuation of hypoxic conditions or after return to oxygenated conditions. Parallel segments of pulmonary artery from normal dogs were also studied. Norepinephrine stores were labeled with [3H]norepinephrine and measurements were made of [3H]norepinephrine and its radiolabeled metabolites (separated by column chromatography) in superfusates using liquid scintillation spectrometry. Chronic hypoxia (1) reduced neuronal uptake of NE from synaptic clefts, (2) reduced the content of DOPEG in superfusate from tissues studied during continuation of hypoxic conditions and in tissues studied after return to oxygenated conditions, (3) increased extraneuronal uptake of NE and (4) increased overflow of NE from synaptic clefts. In similar segments of pulmonary artery removed from the same lung, endogenous free and conjugated norepinephrine and dopamine were measured in pulmonary artery by liquid chromatography with electrochemical detection. The tissue content of free norepinephrine after stimulation was reduced, which was compatible with the reduction in neuronal uptake. Conjugated norepinephrine was a minor metabolite and was increased modestly compared to concentrations reported previously in pulmonary artery from normal dogs.