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Plasma membrane calcium ATPase plays a role in reducing Ca(2+)-mediated cytotoxicity in PC12 cells

M L Garcia1, Y M Usachev, S A Thayer

  • 1Program in Molecular Neuroscience, Mayo Graduate School, Rochester, Minnesota, USA.

Journal of Neuroscience Research
|June 9, 2001
PubMed

Insights

Altering plasma membrane calcium ATPase expression significantly impacts cell survival during calcium overload. Overexpressing this calcium pump enhances cell resistance, while suppressing it increases vulnerability to cell death.

Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Cell death from various insults often involves increased intracellular calcium (Ca2+).
  • Mechanisms of Ca2+ homeostasis affected during insults remain unclear.
  • Plasma membrane calcium ATPase (PMCA) mRNA expression changes in vulnerable neurons during seizures.

Purpose of the Study:

  • To investigate the role of PMCA expression in cellular vulnerability to Ca2+ overload.
  • To determine if modulating PMCA levels affects cell death in a model system.

Main Methods:

  • Utilized rat pheochromocytoma 12 cells, known to be vulnerable to Ca2+ overload.
  • Employed reverse transcriptase-PCR and Western blot to identify PMCA isoforms.
  • Created stable cell lines overexpressing or suppressing PMCA isoform 4b.
  • Assessed cell viability following exposure to the Ca2+ ionophore A23187.

Main Results:

  • PMCA isoform 4b was identified as a major Ca2+ pump in pheochromocytoma 12 cells.
  • Cells overexpressing PMCA 4b showed reduced vulnerability to A23187-induced cell death.
  • Cells with suppressed PMCA 4b expression (antisense clones) were significantly more susceptible to Ca2+-mediated death.

Conclusions:

  • Regulation of PMCA expression is crucial for cellular survival under pathological intracellular calcium conditions.
  • PMCA activity plays a key role in protecting cells from Ca2+ overload-induced death.
  • Modulating PMCA levels offers a potential therapeutic target for conditions involving excitotoxicity and calcium dysregulation.

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