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Originally published In Press as doi:10.1074/jbc.M207199200 on October 29, 2002

J. Biol. Chem., Vol. 278, Issue 1, 200-207, January 3, 2003
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Ca2+ Activates Cystic Fibrosis Transmembrane Conductance Regulator- and Clminus -dependent HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> Transport in Pancreatic Duct Cells*

Wan NamkungDagger , Jin Ah LeeDagger , Wooin AhnDagger , WonSun HanDagger , Sung Won KwonDagger , Duk Sun Ahn§, Kyung Hwan KimDagger , and Min Goo LeeDagger

From the Dagger  Department of Pharmacology and Brain Korea 21 Project for Medical Sciences and § Department of Physiology, Yonsei University College of Medicine, Seoul 120-752, Korea

Pancreatic duct cells secrete bicarbonate-rich fluids, which are important for maintaining the patency of pancreatic ductal trees as well as intestinal digestive function. The bulk of bicarbonate secretion in the luminal membrane of duct cells is mediated by a Cl--dependent mechanism (Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchange), and we previously reported that the mechanism is CFTR-dependent and cAMP-activated (Lee, M. G., Choi, J. Y., Luo, X., Strickland, E., Thomas, P. J., and Muallem, S. (1999) J. Biol. Chem. 274, 14670-14677). In the present study, we provide comprehensive evidence that calcium signaling also activates the same CFTR- and Cl--dependent HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> transport. ATP and trypsin evoked intracellular calcium signaling in pancreatic duct-derived cells through the activation of purinergic and protease-activated receptors, respectively. Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchange activity was measured by recording pHi in response to [Cl-]o changes of the perfusate. In perfusate containing high concentrations of K+, which blocks Cl- movement through electrogenic or K+-coupled pathways, ATP and trypsin highly stimulated luminal Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchange activity in CAPAN-1 cells expressing wild-type CFTR, but not in CFPAC-1 cells that have defective (Delta F508) CFTR. Notably, adenoviral transfection of wild-type CFTR in CFPAC-1 cells completely restored the stimulatory effect of ATP on luminal Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchange. In addition, the chelation of intracellular calcium by 1,2-bis(2-aminophenoxy)ethane-N,N,N,N'-tetraacetic acid (BAPTA) treatment abolished the effect of calcium agonists on luminal Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchange. These results provide a molecular basis for calcium-induced bicarbonate secretion in pancreatic duct cells and highlight the importance of CFTR in epithelial bicarbonate secretion induced by various stimuli.

* This work was supported by grants R02-2002-000-00052-0 from the Basic Research Program of the Korea Science and Engineering Foundation and by HMP-00-B-21400-0059 from the Ministry of Health and Welfare, Korea (to M. G. L.) and from the Brain Korea 21 Project for Medical Sciences, Yonsei University, Korea (to K. H. K.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Department of Pharmacology, Yonsei University College of Medicine, 134 Sinchon-dong, Seoul 120-752, Korea. Tel.: 82-2-361-5221; Fax: 82-2-313-1894; E-mail: mlee@yumc.yonsei.ac.kr.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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