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Plasma membrane voltage-dependent anion channel mediates antiestrogen-activated maxi Cl- currents in C1300

Maria I Bahamonde1, José M Fernández-Fernández, Francesc X Guix

  • 1Unitat de Senyalització Cel.lular, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, C/ Dr. Aiguader 80, Barcelona 08003, Spain.

The Journal of Biological Chemistry
|June 10, 2003
PubMed

Insights

The voltage-dependent anion channel (VDAC) may form the plasma membrane Maxi Chloride channel. This study demonstrates VDAC

Area of Science:

  • Cell Biology
  • Neuroscience
  • Biophysics

Background:

  • The molecular identity of the large conductance voltage-dependent chloride channel (Maxi Cl- channel) in the plasma membrane remains unknown.
  • Electrophysiological properties of the Maxi Cl- channel resemble the voltage-dependent anion channel (VDAC) found in the outer mitochondrial membrane.
  • A controversial hypothesis suggests VDAC might be the molecular basis for the plasma membrane Maxi Cl- channel.

Purpose of the Study:

  • To investigate the cellular localization of VDAC.
  • To determine the relationship between VDAC and antiestrogen-activated Maxi Cl- currents in neuroblastoma cells.
  • To elucidate the molecular nature of the plasma membrane Maxi Cl- channel.

Main Methods:

  • Immunoblotting of membrane fractions using VDAC-specific monoclonal antibodies.
  • Reverse transcription-PCR to detect VDAC mRNA with a specific leader peptide sequence.
  • Immunofluorescence microscopy to assess VDAC colocalization with plasma membrane lipid raft markers.
  • Antisense oligonucleotide transfection to inhibit VDAC expression.

Main Results:

  • Plasma membrane VDAC was detected in C1300 neuroblastoma cells.
  • VDAC colocalized with cholera toxin beta subunit (a lipid raft marker) but not caveolin-1.
  • Inhibition of VDAC expression using antisense oligonucleotides significantly reduced both VDAC immunostaining and Maxi Cl- currents activated by antiestrogens.

Conclusions:

  • The findings suggest that VDAC is present in the plasma membrane of neuroblastoma cells.
  • VDAC likely forms the plasma membrane Maxi Cl- channel or is a component thereof.
  • This research provides evidence supporting the controversial hypothesis linking VDAC to the Maxi Cl- channel.

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