Ca2+ activates cystic fibrosis transmembrane conductance regulator- and Cl--dependent HCO3- transport in pancreatic duct cells

Wan Namkung, Jin Ah Lee, Wooin Ahn, Won Sun Han, Sung Won Kwon, Duk Sun Ahn, Kyung Hwan Kim, Min Goo Lee

Research output: Contribution to journalArticle

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Abstract

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-/HCO3- 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 HCO3- transport. ATP and trypsin evoked intracellular calcium signaling in pancreatic duct-derived cells through the activation of purinergic and protease-activated receptors, respectively. Cl-/HCO3- 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-/HCO3- exchange activity in CAPAN-1 cells expressing wild-type CFTR, but not in CFPAC-1 cells that have defective (ΑF508) CFTR. Notably, adenoviral transfection of wild-type CFTR in CFPAC-1 cells completely restored the stimulatory effect of ATP on luminal Cl-/HCO3- exchange. In addition, the chelation of intracellular calcium by 1,2bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid (BAPTA) treatment abolished the effect of calcium agonists on luminal Cl-/HCO3- 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.

Original languageEnglish
Pages (from-to)200-207
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number1
DOIs
Publication statusPublished - 2003 Jan 3

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Cystic Fibrosis Transmembrane Conductance Regulator
Pancreatic Ducts
Ducts
Bicarbonates
Calcium
Adenosine Triphosphate
Calcium Signaling
Trypsin
Proteinase-Activated Receptors
Ethane
Chelation
Ion exchange
Chemical activation
Transfection
Cells
Membranes
Cell Membrane
Acids
Fluids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Namkung, Wan ; Lee, Jin Ah ; Ahn, Wooin ; Han, Won Sun ; Kwon, Sung Won ; Ahn, Duk Sun ; Kim, Kyung Hwan ; Lee, Min Goo. / Ca2+ activates cystic fibrosis transmembrane conductance regulator- and Cl--dependent HCO3- transport in pancreatic duct cells. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 1. pp. 200-207.
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title = "Ca2+ activates cystic fibrosis transmembrane conductance regulator- and Cl--dependent HCO3- transport in pancreatic duct cells",
abstract = "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-/HCO3- 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 HCO3- transport. ATP and trypsin evoked intracellular calcium signaling in pancreatic duct-derived cells through the activation of purinergic and protease-activated receptors, respectively. Cl-/HCO3- 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-/HCO3- exchange activity in CAPAN-1 cells expressing wild-type CFTR, but not in CFPAC-1 cells that have defective (ΑF508) CFTR. Notably, adenoviral transfection of wild-type CFTR in CFPAC-1 cells completely restored the stimulatory effect of ATP on luminal Cl-/HCO3- exchange. In addition, the chelation of intracellular calcium by 1,2bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid (BAPTA) treatment abolished the effect of calcium agonists on luminal Cl-/HCO3- 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.",
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Ca2+ activates cystic fibrosis transmembrane conductance regulator- and Cl--dependent HCO3- transport in pancreatic duct cells. / Namkung, Wan; Lee, Jin Ah; Ahn, Wooin; Han, Won Sun; Kwon, Sung Won; Ahn, Duk Sun; Kim, Kyung Hwan; Lee, Min Goo.

In: Journal of Biological Chemistry, Vol. 278, No. 1, 03.01.2003, p. 200-207.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Ca2+ activates cystic fibrosis transmembrane conductance regulator- and Cl--dependent HCO3- transport in pancreatic duct cells

AU - Namkung, Wan

AU - Lee, Jin Ah

AU - Ahn, Wooin

AU - Han, Won Sun

AU - Kwon, Sung Won

AU - Ahn, Duk Sun

AU - Kim, Kyung Hwan

AU - Lee, Min Goo

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N2 - 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-/HCO3- 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 HCO3- transport. ATP and trypsin evoked intracellular calcium signaling in pancreatic duct-derived cells through the activation of purinergic and protease-activated receptors, respectively. Cl-/HCO3- 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-/HCO3- exchange activity in CAPAN-1 cells expressing wild-type CFTR, but not in CFPAC-1 cells that have defective (ΑF508) CFTR. Notably, adenoviral transfection of wild-type CFTR in CFPAC-1 cells completely restored the stimulatory effect of ATP on luminal Cl-/HCO3- exchange. In addition, the chelation of intracellular calcium by 1,2bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid (BAPTA) treatment abolished the effect of calcium agonists on luminal Cl-/HCO3- 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.

AB - 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-/HCO3- 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 HCO3- transport. ATP and trypsin evoked intracellular calcium signaling in pancreatic duct-derived cells through the activation of purinergic and protease-activated receptors, respectively. Cl-/HCO3- 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-/HCO3- exchange activity in CAPAN-1 cells expressing wild-type CFTR, but not in CFPAC-1 cells that have defective (ΑF508) CFTR. Notably, adenoviral transfection of wild-type CFTR in CFPAC-1 cells completely restored the stimulatory effect of ATP on luminal Cl-/HCO3- exchange. In addition, the chelation of intracellular calcium by 1,2bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid (BAPTA) treatment abolished the effect of calcium agonists on luminal Cl-/HCO3- 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.

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