Bicarbonate transport by the human pancreatic ductal cell line hpaf

Irma Demeter, Orsolya Hegyesi, Ákos Károly Nagy, Maynard R. Case, Martin C. Steward, G. Varga, B. Burghardt

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

OBJECTIVES:: The human pancreatic duct cell line, HPAF, has been shown previously to secrete Cl in response to Ca-mobilizing stimuli. Our aim was to assess the capacity of HPAF cells to transport and secrete HCO3. METHODS:: HPAF cells were grown as confluent monolayers on permeable supports. Short-circuit current was measured by voltage clamp. Intracellular pH (pHi) was measured by microfluorometry in cells loaded with 2′,7′-bis(2-carboxyethyl)-5(6) -carboxyfluorescein (BCECF). RESULTS:: In HCO3-free solutions, ATP-evoked changes in short-circuit current were inhibited by bumetanide, and the recovery of pHi from acid loading was abolished by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). In the presence of HCO3, ATP-evoked secretion was no longer inhibited by bumetanide, and there was a strong EIPA-insensitive recovery from acid loading, which was inhibited by 4,4′-diisothiocyanatodihydrostilbene-2,2′- disulfonate (H2DIDS). ATP, but not forskolin, stimulated HCO3 efflux from the cells. CONCLUSIONS:: In the absence of HCO3, ATP-evoked Cl secretion is driven by a basolateral Na-K-2Cl cotransporter, and pHi is regulated by apical and basolateral Na/H exchangers. In the presence of HCO3, ATP-evoked secretion is sustained in the absence of Na-K-2Cl cotransporter activity and is probably driven by basolateral Na-HCO3 cotransport.

Original languageEnglish
Pages (from-to)913-920
Number of pages8
JournalPancreas.
Volume38
Issue number8
DOIs
Publication statusPublished - Nov 2009

Fingerprint

Bicarbonates
Adenosine Triphosphate
Cell Line
Bumetanide
Cytophotometry
Sodium-Hydrogen Antiporter
Acids
Amiloride
Pancreatic Ducts
Colforsin

Keywords

  • Bicarbonate transport
  • CaCC
  • CFTR
  • Intracellular pH
  • Pancreatic duct

ASJC Scopus subject areas

  • Hepatology
  • Internal Medicine
  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Bicarbonate transport by the human pancreatic ductal cell line hpaf. / Demeter, Irma; Hegyesi, Orsolya; Nagy, Ákos Károly; Case, Maynard R.; Steward, Martin C.; Varga, G.; Burghardt, B.

In: Pancreas., Vol. 38, No. 8, 11.2009, p. 913-920.

Research output: Contribution to journalArticle

Demeter, I, Hegyesi, O, Nagy, ÁK, Case, MR, Steward, MC, Varga, G & Burghardt, B 2009, 'Bicarbonate transport by the human pancreatic ductal cell line hpaf', Pancreas., vol. 38, no. 8, pp. 913-920. https://doi.org/10.1097/MPA.0b013e3181b32c08
Demeter I, Hegyesi O, Nagy ÁK, Case MR, Steward MC, Varga G et al. Bicarbonate transport by the human pancreatic ductal cell line hpaf. Pancreas. 2009 Nov;38(8):913-920. https://doi.org/10.1097/MPA.0b013e3181b32c08
Demeter, Irma ; Hegyesi, Orsolya ; Nagy, Ákos Károly ; Case, Maynard R. ; Steward, Martin C. ; Varga, G. ; Burghardt, B. / Bicarbonate transport by the human pancreatic ductal cell line hpaf. In: Pancreas. 2009 ; Vol. 38, No. 8. pp. 913-920.
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abstract = "OBJECTIVES:: The human pancreatic duct cell line, HPAF, has been shown previously to secrete Cl in response to Ca-mobilizing stimuli. Our aim was to assess the capacity of HPAF cells to transport and secrete HCO3. METHODS:: HPAF cells were grown as confluent monolayers on permeable supports. Short-circuit current was measured by voltage clamp. Intracellular pH (pHi) was measured by microfluorometry in cells loaded with 2′,7′-bis(2-carboxyethyl)-5(6) -carboxyfluorescein (BCECF). RESULTS:: In HCO3-free solutions, ATP-evoked changes in short-circuit current were inhibited by bumetanide, and the recovery of pHi from acid loading was abolished by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). In the presence of HCO3, ATP-evoked secretion was no longer inhibited by bumetanide, and there was a strong EIPA-insensitive recovery from acid loading, which was inhibited by 4,4′-diisothiocyanatodihydrostilbene-2,2′- disulfonate (H2DIDS). ATP, but not forskolin, stimulated HCO3 efflux from the cells. CONCLUSIONS:: In the absence of HCO3, ATP-evoked Cl secretion is driven by a basolateral Na-K-2Cl cotransporter, and pHi is regulated by apical and basolateral Na/H exchangers. In the presence of HCO3, ATP-evoked secretion is sustained in the absence of Na-K-2Cl cotransporter activity and is probably driven by basolateral Na-HCO3 cotransport.",
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T1 - Bicarbonate transport by the human pancreatic ductal cell line hpaf

AU - Demeter, Irma

AU - Hegyesi, Orsolya

AU - Nagy, Ákos Károly

AU - Case, Maynard R.

AU - Steward, Martin C.

AU - Varga, G.

AU - Burghardt, B.

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N2 - OBJECTIVES:: The human pancreatic duct cell line, HPAF, has been shown previously to secrete Cl in response to Ca-mobilizing stimuli. Our aim was to assess the capacity of HPAF cells to transport and secrete HCO3. METHODS:: HPAF cells were grown as confluent monolayers on permeable supports. Short-circuit current was measured by voltage clamp. Intracellular pH (pHi) was measured by microfluorometry in cells loaded with 2′,7′-bis(2-carboxyethyl)-5(6) -carboxyfluorescein (BCECF). RESULTS:: In HCO3-free solutions, ATP-evoked changes in short-circuit current were inhibited by bumetanide, and the recovery of pHi from acid loading was abolished by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). In the presence of HCO3, ATP-evoked secretion was no longer inhibited by bumetanide, and there was a strong EIPA-insensitive recovery from acid loading, which was inhibited by 4,4′-diisothiocyanatodihydrostilbene-2,2′- disulfonate (H2DIDS). ATP, but not forskolin, stimulated HCO3 efflux from the cells. CONCLUSIONS:: In the absence of HCO3, ATP-evoked Cl secretion is driven by a basolateral Na-K-2Cl cotransporter, and pHi is regulated by apical and basolateral Na/H exchangers. In the presence of HCO3, ATP-evoked secretion is sustained in the absence of Na-K-2Cl cotransporter activity and is probably driven by basolateral Na-HCO3 cotransport.

AB - OBJECTIVES:: The human pancreatic duct cell line, HPAF, has been shown previously to secrete Cl in response to Ca-mobilizing stimuli. Our aim was to assess the capacity of HPAF cells to transport and secrete HCO3. METHODS:: HPAF cells were grown as confluent monolayers on permeable supports. Short-circuit current was measured by voltage clamp. Intracellular pH (pHi) was measured by microfluorometry in cells loaded with 2′,7′-bis(2-carboxyethyl)-5(6) -carboxyfluorescein (BCECF). RESULTS:: In HCO3-free solutions, ATP-evoked changes in short-circuit current were inhibited by bumetanide, and the recovery of pHi from acid loading was abolished by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). In the presence of HCO3, ATP-evoked secretion was no longer inhibited by bumetanide, and there was a strong EIPA-insensitive recovery from acid loading, which was inhibited by 4,4′-diisothiocyanatodihydrostilbene-2,2′- disulfonate (H2DIDS). ATP, but not forskolin, stimulated HCO3 efflux from the cells. CONCLUSIONS:: In the absence of HCO3, ATP-evoked Cl secretion is driven by a basolateral Na-K-2Cl cotransporter, and pHi is regulated by apical and basolateral Na/H exchangers. In the presence of HCO3, ATP-evoked secretion is sustained in the absence of Na-K-2Cl cotransporter activity and is probably driven by basolateral Na-HCO3 cotransport.

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KW - CaCC

KW - CFTR

KW - Intracellular pH

KW - Pancreatic duct

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