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Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line.

Publication ,  Journal Article
Fan, PY; Haas, M; Middleton, JP
Published in: Biochim Biophys Acta
October 19, 1992

Lack of an adequate cell model has limited investigation of Na/K/Cl cotransporter regulation in the kidney. Using A6 cells, an amphibian distal renal cell line, we observed that 63% of rubidium uptake in confluent A6 monolayers was ouabain-insensitive. Ouabain-insensitive rubidium uptake was inhibited in a dose-dependent fashion by furosemide (IC50 6.6 microM) or bumetanide (IC50 1.7 microM). Kinetic studies confirmed that furosemide-sensitive rubidium uptake had features consistent with cotransporter activity in other cell lines. Furthermore, specific binding of [3H]bumetanide occurred with a capacity of 8.6 pmol/mg protein and a Kd of 1.6 microM bumetanide. Finally, furosemide-sensitive rubidium uptake was rapidly regulated by a calcium ionophore, the phorbol ester PDBu, forskolin, and adenosine. These data demonstrate an Na/K/Cl cotransport system in the A6 cell which will serve as a useful model for studying cotransporter regulation by endogenous signaling pathways.

Duke Scholars

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

October 19, 1992

Volume

1111

Issue

1

Start / End Page

75 / 80

Location

Netherlands

Related Subject Headings

  • Xenopus laevis
  • Sodium-Potassium-Exchanging ATPase
  • Sodium
  • Potassium
  • Kidney
  • Ion Transport
  • Furosemide
  • Chlorides
  • Cell Line
  • Animals
 

Citation

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Fan, P. Y., Haas, M., & Middleton, J. P. (1992). Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line. Biochim Biophys Acta, 1111(1), 75–80. https://doi.org/10.1016/0005-2736(92)90276-r
Fan, P. Y., M. Haas, and J. P. Middleton. “Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line.Biochim Biophys Acta 1111, no. 1 (October 19, 1992): 75–80. https://doi.org/10.1016/0005-2736(92)90276-r.
Fan PY, Haas M, Middleton JP. Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line. Biochim Biophys Acta. 1992 Oct 19;1111(1):75–80.
Fan, P. Y., et al. “Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line.Biochim Biophys Acta, vol. 1111, no. 1, Oct. 1992, pp. 75–80. Pubmed, doi:10.1016/0005-2736(92)90276-r.
Fan PY, Haas M, Middleton JP. Identification of a regulated Na/K/Cl cotransport system in a distal nephron cell line. Biochim Biophys Acta. 1992 Oct 19;1111(1):75–80.

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

October 19, 1992

Volume

1111

Issue

1

Start / End Page

75 / 80

Location

Netherlands

Related Subject Headings

  • Xenopus laevis
  • Sodium-Potassium-Exchanging ATPase
  • Sodium
  • Potassium
  • Kidney
  • Ion Transport
  • Furosemide
  • Chlorides
  • Cell Line
  • Animals