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J Appl Physiol 94: 343-352, 2003. First published August 30, 2002; doi:10.1152/japplphysiol.00562.2002
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Vol. 94, Issue 1, 343-352, January 2003

Pleural surface fluorescence measurement of Na+ and Clminus transport across the air space-capillary barrier

Jinjun Jiang1,*, Yuanlin Song1,*, Chunxue Bai2, Beverly H. Koller3, Michael A. Matthay1, and A. S. Verkman1

1 Departments of Medicine and Physiology, Cardiovascular Research Institute, University of California, San Francisco, California 94143 - 0521; 2 Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai, China; and 3 Cystic Fibrosis/Pulmonary Research Center, University of North Carolina, Chapel Hill, North Carolina 37599 - 7248

We developed a pleural surface fluorescence method to measure Na+ and Cl- transport in perfused mouse lungs. The air space was filled with aqueous fluid containing membrane-impermeant fluorescent indicators of Cl- (lucigenin) or Na+ (Sodium Green). After instillation of a Cl--free solution into the air space, an increase in perfusate Cl- concentration from 0 to 30 mM produced a decrease in surface lucigenin fluorescence (6.5%/min) corresponding to Cl- influx of 1.0 mM/min. Cl- influx was increased to 2.1 ± 0.3 mM/min by forskolin, and the increase was inhibited by glibenclamide. cAMP-stimulated Cl- influx was decreased by 57% in CFTR null mice. After instillation of a Na+-free solution into the air space, an increase in perfusate Na+ concentration from 0 to 30 mM gave increased Sodium Green fluorescence (Na+ influx of 1.2 mM/min), which increased approximately fivefold after cAMP agonists. Cl- and Na+ transport were not affected in lungs from mice lacking aquaporins AQP1 or AQP5. Our results establish a pleural surface fluorescence method to measure unidirectional Cl- and Na+ flux in intact lung and provide evidence for cAMP-stimulated transcellular Cl- and Na+ transport.

fluorescent indicators; alveolus; lung perfusion; cystic fibrosis transmembrane conductance regulator; ion transport; aquaporin

* J. Jiang and Y. Song contributed equally to this work.




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