High Microvascular Endothelial Water Permeability in Mouse Lung Measured by a Pleural Surface Fluorescence Method

Abstract

Transport of water between the capillary and airspace compartments in lung encounters serial barriers: the alveolar epithelium, interstitium, and capillary endothelium. We previously reported a pleural surface fluorescence method to measure net capillary-to-airspace water transport. To measure the osmotic water permeability across the microvascular endothelial barrier in intact lung, the airspace was filled with a water-immiscible fluorocarbon. The capillaries were perfused via the pulmonary artery with solutions of specified osmolalites containing a high-molecular-weight fluorescent dextran. An increase in perfusate osmolality produced a prompt decrease in surface fluorescence due to dye dilution in the capillaries, followed by a slower return to initial fluorescence as capillary and lung interstitial osmolality equilibrate. A mathematical model was developed to determine the osmotic water permeability coefficient ( P f) of lung microvessels from the time course of pleural surface fluorescence. As predicted, the magnitude of the prompt change in surface fluorescence increased with decreased pulmonary artery perfusion rate and increased osmotic gradient size. With raffinose used to induce the osmotic gradient, P f was 0.03 cm/s at 23°C and was reduced 54% by 0.5 mM HgCl 2. Temperature dependence measurements gave an Arrhenius activation energy ( E a) of 5.4 kcal/mol (12–37°C). The apparent P f induced by the smaller osmolytes mannitol and glycine was 0.021 and 0.011 cm/s (23°C). Immunoblot analysis showed ∼1.4 × 10 12 aquaporin-1 water channels/cm 2 of capillary surface, which accounted quantitatively for the high P f. These results establish a novel method for measuring osmotically driven water permeability across microvessels in intact lung. The high P f, low E a, and mercurial inhibition indicate the involvement of molecular water channels in water transport across the lung endothelium.