Electrophysiological characterization of volume-activated chloride currents in mouse cholangiocyte cell line

Biyi Chen, Grant Nicol, Won Kyoo Cho

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9 Citations (Scopus)

Abstract

Recent electrophysiological and radioisotope efflux studies have demonstrated various Cl- channels in cholangiocytes including volume-activated Cl- channels (VACC). Because VACCs play prominent roles in many vital cellular functions and physiology in cholangiocytes, we have examined their electrophysiological characteristics in mouse cholangiocytes to provide an important framework for studying in the future. The present study is to characterize VACCs expressed in the mouse bile duct cell (MBDC) line, conditionally immortalized by SV40 virus. Conventional whole cell patch-clamp techniques were used to study the electrophysiological characteristics of VACC in MBDC. When the MBDCs were exposed to hypotonic solution, they exhibited an outwardly rectified current, which was significantly inhibited by replacing chloride in the bath solution with gluconate or glutamate and by administration of classic chloride channel inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoate, glybenclamide, DIDS, and tamoxifen. These inhibitory effects were reversible with washing them out from the bath solution. Moreover, the ion selectivity of the volume-activated channel to different anions indicates that it is more permeable to SCN- > I- ≥ Cl- > F - ≥ acetate ≥ glutamate ≥ gluconate. These electrophysiological characteristics demonstrate that the volume-activated current observed is a VACC. In addition, the VACC in MBDC has electrophysiological characteristics similar to those of the VACC in human cholangiocarcinoma cell line. The present study is the first to characterize the VACC in mouse cholangiocyte and will provide an important framework for further studies to examine and understand the role of the VACC in biliary secretion and ion-transport physiology.

Original languageEnglish
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume287
Issue number6 50-6
DOIs
StatePublished - Dec 2004

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Chlorides
Cell Line
Bile Ducts
Baths
Glutamic Acid
Hypotonic Solutions
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Simian virus 40
Cholangiocarcinoma
Glyburide
Benzoates
Ion Transport
Patch-Clamp Techniques
Tamoxifen
Radioisotopes
Anions
Acetates
Ions
gluconic acid

Keywords

  • Bile duct cell
  • Cell volume
  • Ion channel
  • Mouse cholangiocyte
  • Patch clamping
  • Regulatory volume decrease
  • Volume-activated chloride channel

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology

Cite this

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title = "Electrophysiological characterization of volume-activated chloride currents in mouse cholangiocyte cell line",
abstract = "Recent electrophysiological and radioisotope efflux studies have demonstrated various Cl- channels in cholangiocytes including volume-activated Cl- channels (VACC). Because VACCs play prominent roles in many vital cellular functions and physiology in cholangiocytes, we have examined their electrophysiological characteristics in mouse cholangiocytes to provide an important framework for studying in the future. The present study is to characterize VACCs expressed in the mouse bile duct cell (MBDC) line, conditionally immortalized by SV40 virus. Conventional whole cell patch-clamp techniques were used to study the electrophysiological characteristics of VACC in MBDC. When the MBDCs were exposed to hypotonic solution, they exhibited an outwardly rectified current, which was significantly inhibited by replacing chloride in the bath solution with gluconate or glutamate and by administration of classic chloride channel inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoate, glybenclamide, DIDS, and tamoxifen. These inhibitory effects were reversible with washing them out from the bath solution. Moreover, the ion selectivity of the volume-activated channel to different anions indicates that it is more permeable to SCN- > I- ≥ Cl- > F - ≥ acetate ≥ glutamate ≥ gluconate. These electrophysiological characteristics demonstrate that the volume-activated current observed is a VACC. In addition, the VACC in MBDC has electrophysiological characteristics similar to those of the VACC in human cholangiocarcinoma cell line. The present study is the first to characterize the VACC in mouse cholangiocyte and will provide an important framework for further studies to examine and understand the role of the VACC in biliary secretion and ion-transport physiology.",
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T1 - Electrophysiological characterization of volume-activated chloride currents in mouse cholangiocyte cell line

AU - Chen, Biyi

AU - Nicol, Grant

AU - Cho, Won Kyoo

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N2 - Recent electrophysiological and radioisotope efflux studies have demonstrated various Cl- channels in cholangiocytes including volume-activated Cl- channels (VACC). Because VACCs play prominent roles in many vital cellular functions and physiology in cholangiocytes, we have examined their electrophysiological characteristics in mouse cholangiocytes to provide an important framework for studying in the future. The present study is to characterize VACCs expressed in the mouse bile duct cell (MBDC) line, conditionally immortalized by SV40 virus. Conventional whole cell patch-clamp techniques were used to study the electrophysiological characteristics of VACC in MBDC. When the MBDCs were exposed to hypotonic solution, they exhibited an outwardly rectified current, which was significantly inhibited by replacing chloride in the bath solution with gluconate or glutamate and by administration of classic chloride channel inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoate, glybenclamide, DIDS, and tamoxifen. These inhibitory effects were reversible with washing them out from the bath solution. Moreover, the ion selectivity of the volume-activated channel to different anions indicates that it is more permeable to SCN- > I- ≥ Cl- > F - ≥ acetate ≥ glutamate ≥ gluconate. These electrophysiological characteristics demonstrate that the volume-activated current observed is a VACC. In addition, the VACC in MBDC has electrophysiological characteristics similar to those of the VACC in human cholangiocarcinoma cell line. The present study is the first to characterize the VACC in mouse cholangiocyte and will provide an important framework for further studies to examine and understand the role of the VACC in biliary secretion and ion-transport physiology.

AB - Recent electrophysiological and radioisotope efflux studies have demonstrated various Cl- channels in cholangiocytes including volume-activated Cl- channels (VACC). Because VACCs play prominent roles in many vital cellular functions and physiology in cholangiocytes, we have examined their electrophysiological characteristics in mouse cholangiocytes to provide an important framework for studying in the future. The present study is to characterize VACCs expressed in the mouse bile duct cell (MBDC) line, conditionally immortalized by SV40 virus. Conventional whole cell patch-clamp techniques were used to study the electrophysiological characteristics of VACC in MBDC. When the MBDCs were exposed to hypotonic solution, they exhibited an outwardly rectified current, which was significantly inhibited by replacing chloride in the bath solution with gluconate or glutamate and by administration of classic chloride channel inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoate, glybenclamide, DIDS, and tamoxifen. These inhibitory effects were reversible with washing them out from the bath solution. Moreover, the ion selectivity of the volume-activated channel to different anions indicates that it is more permeable to SCN- > I- ≥ Cl- > F - ≥ acetate ≥ glutamate ≥ gluconate. These electrophysiological characteristics demonstrate that the volume-activated current observed is a VACC. In addition, the VACC in MBDC has electrophysiological characteristics similar to those of the VACC in human cholangiocarcinoma cell line. The present study is the first to characterize the VACC in mouse cholangiocyte and will provide an important framework for further studies to examine and understand the role of the VACC in biliary secretion and ion-transport physiology.

KW - Bile duct cell

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KW - Patch clamping

KW - Regulatory volume decrease

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