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Articles in PresS, published online ahead of print August 30, 2002
J Appl Physiol, 10.1152/jap.00562.2002
Submitted on June 27, 2002
Accepted on August 23, 2002
1 Departments of Medicine and Physiology, University of California, San Francisco, San Francisco, CA, USA
2 Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai, China
3 Cystic Fibrosis/Pulmonary Research Center, University of North Carolina, Chapel Hill, NC, USA
* To whom correspondence should be addressed. E-mail: verkman{at}itsa.ucsf.edu.
We developed a pleural surface fluorescence method to measure Na+ and Cl- transport in perfused mouse lungs. The airspace was filled with aqueous fluid containing membrane-impermeant fluorescent indicators of Cl- (lucigenin) or Na+ (Sodium-greenTM). After instillation of a Cl- free solution into the airspace, an increase in perfusate [Cl-] 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 airspace, an increase in perfusate [Na+] from 0 to 30 mM gave increased Sodium-greenTM fluorescence (Na+ influx 1.2 mM/min), which increased ~5-fold 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.
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