Surfactant phospholipid metabolism and lung stability were studied in mechanically ventilated and in spontaneously breathing rabbits (control group). During ventilation the dynamic lung-thorax compliance decreased to 79% after 6 h. Static compliance and amount and composition of surfactant phospholipids remained unaltered. These data indicate inactivation of alveolar surfactant during ventilation, which is reversible by a single large inflation. Incorporation of injected radioactively labeled palmitate into saturated phosphatidylcholine (SPC) of the lamellar body fraction increased significantly in the ventilated group; maximal specific activity increased from 20 dpm/nmol SPC at 6 h in the control group to 30 dpm/nmol SPC at 2 h in the ventilated group. The clearance of radioactivity from the lamellar bodies into the alveolar lumen was greatly enhanced in the ventilated group. The results are explained by assuming that as a result of the inactivation of alveolar surfactant, endocytosed surfactant is degraded in the type II cells instead of being recycled and that the degradation products are subsequently reutilized in surfactant synthesis. This interpretation is supported by computer simulations.