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Related Experiment Videos

Relationship between Na(+),K(+)-ATPase and cell attachment.

R G Contreras1, L Shoshani, C Flores-Maldonado

  • 1Center for Research and Advanced Studies of México (CINVESTAV), Av. Instituto Politecnico Nacional 2508, Codigo Postal, 07660 Mexico.

Journal of Cell Science
|November 24, 1999
PubMed
Summary
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Ouabain blockade of the sodium-potassium pump (Na(+),K(+)-ATPase) triggers cell detachment by increasing tyrosine phosphorylation and activating MAP kinase signaling pathways. This pump-to-attachment (P-A) mechanism is crucial for cell adhesion.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • The Na(+),K(+)-ATPase (sodium-potassium pump) is vital for cellular homeostasis.
  • Previous observations suggest a link between pump activity and cell attachment.
  • The precise mechanism linking pump function to cell adhesion remains unclear.

Purpose of the Study:

  • To investigate the molecular mechanisms by which ouabain blockade of the Na(+),K(+)-ATPase affects cell adhesion.
  • To identify signaling pathways involved in ouabain-induced cell detachment.
  • To explore the role of specific cell types in this pump-to-attachment (P-A) phenomenon.

Main Methods:

  • Treatment of Madin-Darby Canine Kidney-W (MDCK-W) cells with ouabain.
  • Analysis of tyrosine phosphorylation and MAP kinase activation.

Related Experiment Videos

  • Assessment of cell adhesion molecule distribution (occludin, ZO-1, desmoplakin, etc.).
  • Inhibition studies using genistein (tyrosine kinase inhibitor) and UO126 (MAP kinase kinase inhibitor).
  • Comparison with MDCK cells having low ouabain affinity and Ma104 cells.

    • Main Results:

    • Ouabain treatment induced cell detachment, increased tyrosine phosphorylation, and activated MAP kinase in MDCK-W cells.
    • Key cell attachment molecules were redistributed upon ouabain exposure.
    • Inhibitors of tyrosine kinase and MAP kinase kinase blocked ouabain-induced detachment.
    • Ouabain increased P190(Rho-GAP) content, implicating Rho/Rac and MAPK pathways.
    • MDCK cells with low ouabain affinity and Ma104 cells did not exhibit detachment or associated molecular changes.

    Conclusions:

    • A pump-to-attachment (P-A) mechanism exists, linking Na(+),K(+)-ATPase activity to cell detachment.
    • MAPK and Rho/Rac signaling pathways are involved in mediating this effect.
    • Cellular differences in Na(+),K(+)-ATPase ouabain affinity influence the response.
    • Ma104 cells appear faulty in transducing the pump blockade signal for detachment.