Cell Biology and Toxicology

, Volume 26, Issue 3, pp 201–213 | Cite as

Diverse actions of ouabain and its aglycone ouabagenin in renal cells

  • Raphael C. Valente
  • Luiz S. Capella
  • Márcia M. M. Oliveira
  • Luciana T. Nunes-Lima
  • Fernanda C. M. Cruz
  • Roberta R. Palmieri
  • Aníbal G. Lopes
  • Márcia A. M. Capella
Original Research


The cellular actions of ouabain are complex and involve different pathways, depending on the cell type and experimental conditions. Several studies have reported that Madin–Darby canine kidney (MDCK) cellular sensitivity to ouabain is not related to Na-K-ATPase inhibition, and others showed that some cell types, such as Ma104, are resistant to ouabain toxicity albeit their Na-K-ATPase isoforms possess high affinity for this glycoside. We describe here that the effects of ouabain and ouabagenin also diverge in MDCK and Ma104 cells, being MDCK cells more resistant to ouabagenin, while Ma104 cells are resistant to both molecules. This feature seems to correlate with induction of cell signaling, since ouabain, but not ouabagenin, induced an intense and sustained increase in tyrosine phosphorylation levels in MDCK cells. Moreover, ouabain-induced phosphorylation in Ma104 cells was approximately half than that observed in MDCK cells. The proportion between α and β subunits of Na-K-ATPase was similar in MDCK cells, though Ma104 cells presented more α subunits, located mainly at the cytoplasm. Furthermore, a fluorescent ouabain-analog labeled mainly the cytoplasm of Ma104 cells, the opposite of that seen in MDCK cells, corroborating the results using anti-Na-K-ATPase antibodies. Hence, the results suggest that ouabain and ouabagenin differ in terms of Na-K-ATPase inhibition and cell signaling activation in MDCK cells. Additionally, MDCK and Ma104 cell lines respond differently to ouabain, perhaps due to an intrinsic ability of this glycoside to selectively reach the cytoplasm of Ma104 cells.


Ouabain Ouabagenin Na-K-ATPase MDCK Ma104 









Phosphorylated tyrosine



This work was supported by grants from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Programa de Oncobiologia (FECD/FAF/ONCO II), PRONEX, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). The authors are indebted to Dr. Elizabeth Giestal de Araujo for the kind donation of monodansylcadaverine.


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Raphael C. Valente
    • 1
  • Luiz S. Capella
    • 2
  • Márcia M. M. Oliveira
    • 2
  • Luciana T. Nunes-Lima
    • 2
  • Fernanda C. M. Cruz
    • 2
  • Roberta R. Palmieri
    • 2
  • Aníbal G. Lopes
    • 2
  • Márcia A. M. Capella
    • 1
    • 2
  1. 1.Instituto de Bioquímica MédicaUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  2. 2.Instituto de Biofísica Carlos Chagas FilhoUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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