Response of sodium pump to ouabain challenge in human glioblastoma cells in culture

World J Biol Psychiatry. 2009;10(4 Pt 3):884-92. doi: 10.1080/15622970902995620.

Abstract

Bipolar disorder is a severe psychiatric condition that manifests with abnormalities in ion regulation. Previous studies have suggested that glia may be specifically involved in the pathophysiology of this condition. Since the potent sodium pump inhibitor, ouabain, has been used previously to model the ionic changes of bipolar illness, we investigated its effect of on sodium pump expression and activity in a human glioblastoma cell line. LN229 cells were grown with or without ouabain 10(-7) M for 3 days, and the effect of a therapeutic concentration of lithium was also examined. The mRNA transcription of sodium pump isoforms was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and the protein expression of phosphorylated and non-phosphorylated pump isoforms was semi-quantified utilizing Western blot. Ouabain treatment caused an increase of some 6-fold in alpha1 protein expression and a doubling of alpha1 mRNA. alpha3 protein and alpha2 and alpha3 mRNA more than doubled. Lithium treatment alone had no effect, but lithium co-administered with ouabain normalized Na pump protein and mRNA expression for alpha1 and 2, but not alpha3. These results suggest that disturbance of ion regulation induces changes in glial cell sodium regulatory systems which are normalized by lithium treatment.

MeSH terms

  • Antipsychotic Agents / therapeutic use
  • Bipolar Disorder / drug therapy
  • Bipolar Disorder / physiopathology*
  • Blotting, Western
  • Brain Neoplasms / genetics
  • Brain Neoplasms / pathology*
  • Cell Line, Tumor / drug effects
  • Cell Line, Tumor / pathology
  • DNA Primers / genetics
  • DNA, Complementary / genetics
  • Enzyme Inhibitors / pharmacology*
  • Enzyme Inhibitors / therapeutic use
  • Glioblastoma / genetics
  • Glioblastoma / pathology*
  • Humans
  • Ion Transport / drug effects
  • Lithium Carbonate / therapeutic use
  • Neuroglia / drug effects*
  • Neuroglia / physiology*
  • Nucleic Acid Amplification Techniques
  • Ouabain / pharmacology*
  • Ouabain / therapeutic use
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sodium-Potassium-Exchanging ATPase / drug effects*
  • Subcellular Fractions

Substances

  • Antipsychotic Agents
  • DNA Primers
  • DNA, Complementary
  • Enzyme Inhibitors
  • RNA, Messenger
  • Lithium Carbonate
  • Ouabain
  • Sodium-Potassium-Exchanging ATPase