The pathogenesis of pulmonary hypertension (PH) remains poorly understood. Vasoconstriction, although likely to be a major factor in the disease, varies between patients and studies of a variety of vasoactive substances have sometimes yielded conflicting results. Amongst these substances, alteration of the nitric oxide (NO) system has been cited as a possible pathogenic factor but both reduction and elevation of the expression of endothelial NO-synthase (eNOS) have been reported in pulmonary vessels. The present study has used immunocytochemistry with well-characterized antibodies to eNOS to investigate its expression in lung tissue taken at transplantation from 44 patients with PH (22 primary, 22 secondary) and 12 non-hypertensive controls. Semi-quantitative assessment showed that although the levels of eNOS expression in pulmonary vessels were variable within both hypertensives and controls, a statistically significant (P<0.01) reduction of immunoreactivity was found in small arterioles from hypertensives compared with controls. In contrast, consistently strong expression of eNOS was seen in the endothelium of plexiform lesions in both the primary and the secondary PH patients. Although a decrease in the NO system of patients with PH has been reported, these findings show a distinct regional distribution of the enzyme with particularly high levels in plexiform lesions, a previously unreported observation, and offer a new perspective on the disease and on the evaluation of possible novel therapeutic approaches.