Intracellular pH regulatory mechanism in a human renal proximal cell line (HKC-8): Evidence for Na+/H+ exchanger, Cl-/HCO3- exchanger and Na+-HCO3- cotransporter

Chiaki Hara, Hiroaki Satoh, Tomohiko Usui, Motoei Kunimi, Eisei Noiri, Kazuhisa Tsukamoto, Shigeo Taniguchi, Shu Uwatoko, Astuo Goto, Lorraine C Racusen, Jun Inatomi, Hitoshi Endou, Toshiro Fujita, George Seki

Research output: Contribution to journalArticle

Abstract

In the present study we investigated whether an immortalized human renal proximal cell line, HKC-8, expresses a recently cloned Na+-HCO3- cotransporter (NBC-1) and, if so, which isoform (kNBC-1 from kidney or pNBC-1 from pancreas) is expressed in this cell line. Cell pH (pH(i)) measurements using a pH-sensitive fluorescence probe in the absence of HCO3-/CO2 revealed the presence of a Na+/H+ exchanger that required high concentrations of amiloride for full inhibition. In the presence of HCO3-/CO2 another pH(i) recovery process, dependent on Na+ but independent of Cl-, was identified. This process was electrogenic and was inhibited by 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), being consistent with the Na+-HCO3- cotransporter. In addition, the pH(i) responses to Cl- removal were compatible with the presence of a Na+-independent Cl-/HCO3- exchanger that was also inhibited by DIDS. Reverse transcriptase polymerase chain reaction (RT-PCR) using primers designed for specific and common regions detected mRNAs of both kNBC-1 and pNBC-1 and Western blot analysis confirmed the expression of NBC-1 protein. These results indicate that HKC-8 has transport activities similar to intact proximal tubules and also suggest that both kNBC-1 and pNBC-1 may contribute to the Na+-HCO3- cotransport activity in this cell line.

Original languageEnglish (US)
Pages (from-to)713-720
Number of pages8
JournalPflugers Archiv European Journal of Physiology
Volume440
Issue number5
StatePublished - 2000

Fingerprint

Chloride-Bicarbonate Antiporters
Sodium-Hydrogen Antiporter
Cells
Kidney
Cell Line
Acids
Amiloride
Polymerase chain reaction
RNA-Directed DNA Polymerase
Protein Isoforms
Fluorescence
Recovery
Messenger RNA
Reverse Transcriptase Polymerase Chain Reaction
Pancreas
Western Blotting
Proteins

Keywords

  • BC-1
  • Cell pH
  • Cl/HCO exchanger
  • HKC-8
  • Na/H exchanger
  • Proximal tubular cells

ASJC Scopus subject areas

  • Physiology

Cite this

Intracellular pH regulatory mechanism in a human renal proximal cell line (HKC-8) : Evidence for Na+/H+ exchanger, Cl-/HCO3- exchanger and Na+-HCO3- cotransporter. / Hara, Chiaki; Satoh, Hiroaki; Usui, Tomohiko; Kunimi, Motoei; Noiri, Eisei; Tsukamoto, Kazuhisa; Taniguchi, Shigeo; Uwatoko, Shu; Goto, Astuo; Racusen, Lorraine C; Inatomi, Jun; Endou, Hitoshi; Fujita, Toshiro; Seki, George.

In: Pflugers Archiv European Journal of Physiology, Vol. 440, No. 5, 2000, p. 713-720.

Research output: Contribution to journalArticle

Hara, C, Satoh, H, Usui, T, Kunimi, M, Noiri, E, Tsukamoto, K, Taniguchi, S, Uwatoko, S, Goto, A, Racusen, LC, Inatomi, J, Endou, H, Fujita, T & Seki, G 2000, 'Intracellular pH regulatory mechanism in a human renal proximal cell line (HKC-8): Evidence for Na+/H+ exchanger, Cl-/HCO3- exchanger and Na+-HCO3- cotransporter', Pflugers Archiv European Journal of Physiology, vol. 440, no. 5, pp. 713-720.
Hara, Chiaki ; Satoh, Hiroaki ; Usui, Tomohiko ; Kunimi, Motoei ; Noiri, Eisei ; Tsukamoto, Kazuhisa ; Taniguchi, Shigeo ; Uwatoko, Shu ; Goto, Astuo ; Racusen, Lorraine C ; Inatomi, Jun ; Endou, Hitoshi ; Fujita, Toshiro ; Seki, George. / Intracellular pH regulatory mechanism in a human renal proximal cell line (HKC-8) : Evidence for Na+/H+ exchanger, Cl-/HCO3- exchanger and Na+-HCO3- cotransporter. In: Pflugers Archiv European Journal of Physiology. 2000 ; Vol. 440, No. 5. pp. 713-720.
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abstract = "In the present study we investigated whether an immortalized human renal proximal cell line, HKC-8, expresses a recently cloned Na+-HCO3- cotransporter (NBC-1) and, if so, which isoform (kNBC-1 from kidney or pNBC-1 from pancreas) is expressed in this cell line. Cell pH (pH(i)) measurements using a pH-sensitive fluorescence probe in the absence of HCO3-/CO2 revealed the presence of a Na+/H+ exchanger that required high concentrations of amiloride for full inhibition. In the presence of HCO3-/CO2 another pH(i) recovery process, dependent on Na+ but independent of Cl-, was identified. This process was electrogenic and was inhibited by 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), being consistent with the Na+-HCO3- cotransporter. In addition, the pH(i) responses to Cl- removal were compatible with the presence of a Na+-independent Cl-/HCO3- exchanger that was also inhibited by DIDS. Reverse transcriptase polymerase chain reaction (RT-PCR) using primers designed for specific and common regions detected mRNAs of both kNBC-1 and pNBC-1 and Western blot analysis confirmed the expression of NBC-1 protein. These results indicate that HKC-8 has transport activities similar to intact proximal tubules and also suggest that both kNBC-1 and pNBC-1 may contribute to the Na+-HCO3- cotransport activity in this cell line.",
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T2 - Evidence for Na+/H+ exchanger, Cl-/HCO3- exchanger and Na+-HCO3- cotransporter

AU - Hara, Chiaki

AU - Satoh, Hiroaki

AU - Usui, Tomohiko

AU - Kunimi, Motoei

AU - Noiri, Eisei

AU - Tsukamoto, Kazuhisa

AU - Taniguchi, Shigeo

AU - Uwatoko, Shu

AU - Goto, Astuo

AU - Racusen, Lorraine C

AU - Inatomi, Jun

AU - Endou, Hitoshi

AU - Fujita, Toshiro

AU - Seki, George

PY - 2000

Y1 - 2000

N2 - In the present study we investigated whether an immortalized human renal proximal cell line, HKC-8, expresses a recently cloned Na+-HCO3- cotransporter (NBC-1) and, if so, which isoform (kNBC-1 from kidney or pNBC-1 from pancreas) is expressed in this cell line. Cell pH (pH(i)) measurements using a pH-sensitive fluorescence probe in the absence of HCO3-/CO2 revealed the presence of a Na+/H+ exchanger that required high concentrations of amiloride for full inhibition. In the presence of HCO3-/CO2 another pH(i) recovery process, dependent on Na+ but independent of Cl-, was identified. This process was electrogenic and was inhibited by 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), being consistent with the Na+-HCO3- cotransporter. In addition, the pH(i) responses to Cl- removal were compatible with the presence of a Na+-independent Cl-/HCO3- exchanger that was also inhibited by DIDS. Reverse transcriptase polymerase chain reaction (RT-PCR) using primers designed for specific and common regions detected mRNAs of both kNBC-1 and pNBC-1 and Western blot analysis confirmed the expression of NBC-1 protein. These results indicate that HKC-8 has transport activities similar to intact proximal tubules and also suggest that both kNBC-1 and pNBC-1 may contribute to the Na+-HCO3- cotransport activity in this cell line.

AB - In the present study we investigated whether an immortalized human renal proximal cell line, HKC-8, expresses a recently cloned Na+-HCO3- cotransporter (NBC-1) and, if so, which isoform (kNBC-1 from kidney or pNBC-1 from pancreas) is expressed in this cell line. Cell pH (pH(i)) measurements using a pH-sensitive fluorescence probe in the absence of HCO3-/CO2 revealed the presence of a Na+/H+ exchanger that required high concentrations of amiloride for full inhibition. In the presence of HCO3-/CO2 another pH(i) recovery process, dependent on Na+ but independent of Cl-, was identified. This process was electrogenic and was inhibited by 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), being consistent with the Na+-HCO3- cotransporter. In addition, the pH(i) responses to Cl- removal were compatible with the presence of a Na+-independent Cl-/HCO3- exchanger that was also inhibited by DIDS. Reverse transcriptase polymerase chain reaction (RT-PCR) using primers designed for specific and common regions detected mRNAs of both kNBC-1 and pNBC-1 and Western blot analysis confirmed the expression of NBC-1 protein. These results indicate that HKC-8 has transport activities similar to intact proximal tubules and also suggest that both kNBC-1 and pNBC-1 may contribute to the Na+-HCO3- cotransport activity in this cell line.

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KW - Na/H exchanger

KW - Proximal tubular cells

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