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Modulation of NMDA-mediated excitotoxicity by protein kinase C

R Wagey1, J Hu, S L Pelech

  • 1Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

Journal of Neurochemistry
|August 25, 2001
PubMed

Insights

Protein kinase C (PKC) activation enhances N-methyl-D-aspartate (NMDA) receptor-mediated cell death in human embryonic kidney cells. This neurotoxicity is specifically mediated by classical PKC isoforms, particularly PKCbeta1.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Cell Biology

Background:

  • Excessive N-methyl-D-aspartate (NMDA) receptor activation causes cell death.
  • Protein kinase C (PKC) is implicated in cellular signaling pathways.

Purpose of the Study:

  • To investigate the role of PKC activation in NMDA receptor-mediated toxicity.
  • To identify the specific PKC isoforms involved in this process.

Main Methods:

  • Utilized human embryonic kidney-293 (HEK) cells transfected with NMDA receptor subunits (NR1/NR2A).
  • Assessed cell survival using trypan blue exclusion assay.
  • Employed phorbol esters, PKC inhibitors, and isoform-specific inhibitors.
  • Measured PKC activity via in vitro kinase assay.

Main Results:

  • NMDA-induced HEK cell death was potentiated by phorbol ester exposure.
  • PKC activation inhibitors or down-regulation reduced NMDA-mediated toxicity.
  • The C-terminus of the NR2A subunit was identified as crucial for PKC-induced enhancement of cell death.
  • Classical PKC isoforms (cPKCs), especially PKCbeta1, mediated the increased cell death.
  • NMDA stimulation increased PKC activity in transfected cells.

Conclusions:

  • PKC activation, particularly via cPKC isoforms like PKCbeta1, accentuates NMDA receptor-mediated neurotoxicity.
  • PKC acts on the NR2A C-terminus to increase cell death.
  • Elevated cellular PKC activity can exacerbate NMDA receptor-induced neurotoxicity.

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