Hypothermia is intentionally imposed during the harvesting of lungs for transplantation. The aim of this study was to investigate the fluid balance alterations in rat lung preparations exposed to hypothermic perfusion. Lowering perfusate temperature from 37 degrees C to values between 27 and 7 degrees C caused an immediate, marked pulmonary hypertension and vasoconstriction accompanied by rapid development of pulmonary edema (+1.15 g, or approximately 90%, gain in lung weight within 5 min). However, on rewarming, vasoconstriction was immediately reversed. Edema was resolved, but along a two-component time course: an immediate reduction of lung weight on rewarming (t 1/2 of 0.5 min) that mirrored the recovery of pulmonary artery pressure and vasoconstriction, and also a slower pressure-independent component of recovery (t 1/2 of 3.5 min). Ouabain (300 microM) markedly inhibited the lung's ability to recover from edema, indicating that fluid clearance from lung tissue was the result of activation of ouabain-sensitive (Na+,K+)-ATPase pump. Results could not be explained by vascular or airspace injury as lung sections from hypothermic lungs appeared normal. The findings indicate that hypothermia induces pulmonary edema formation, which can be rapidly cleared upon rewarming by activation of ouabain-sensitive (Na+,K+)-ATPase pump. Thus, impaired fluid clearance from lung extravascular spaces may be a critical factor limiting gas exchange in transplanted lungs exposed to hypothermia.