Activation of p42 mitogen-activated protein kinase by glutamate receptor stimulation in rat primary cortical cultures

R. S. Fiore, T. H. Murphy, J. S. Sanghera, S. L. Pelech, Jay M Baraban

Research output: Contribution to journalArticle

Abstract

Recent studies have identified at least two homologous mitogen-activated protein (MAP) kinases that are activated by phosphorylation of both tyrosine and threonine residues by an activator kinase. To help define the role of these MAP kinases in neuronal signalling, we have used primary cultures derived from fetal rat cortex to assess the regulation of their activity by agonist stimulation of glutamate receptors and by synaptic activity. Regulation was assayed by monitoring changes in both tyrosine phosphorylation on western blots and in vitro kinase activity toward a selective MAP kinase substrate peptide. In initial studies, we found that phorbol ester treatment increased tyrosine phosphorylation of p42 MAP kinase and stimulated MAP kinase activity. A similar response was elicited by three agonists of metabotropic glutamate receptors, i.e., trans-(±)-1-amino-1,3-cyclopentane dicarboxylic acid, quisqualate, and (2S,3S,4S)-α-(carboxycyclopropyl)glycine. MAP kinase activity and p42 MAP kinase tyrosine phosphorylation were also stimulated by the ionotropic glutamate receptor agonist, kainate, but not by N-methyl-D-aspartate. To examine regulation of MAP kinase by synaptic activity, cultures were treated with picrotoxin, an inhibitor of GABAA receptor-mediated inhibition that enhances spontaneous excitatory synaptic activity. Treatment of cultures with picrotoxin elicited activation of MAP kinase. This response was blocked by tetrodotoxin, which suppresses synaptic activity. These results demonstrate that p42 MAP kinase is activated by glutamate receptor agonist stimulation and by endogenous synaptic activity.

Original languageEnglish (US)
Pages (from-to)1626-1633
Number of pages8
JournalJournal of Neurochemistry
Volume61
Issue number5
StatePublished - Nov 1993

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Mitogen-Activated Protein Kinase 1
Glutamate Receptors
Mitogen-Activated Protein Kinases
Rats
Chemical activation
Phosphorylation
Tyrosine
Excitatory Amino Acid Agonists
Picrotoxin
Phosphotransferases
Quisqualic Acid
Ionotropic Glutamate Receptors
Cyclopentanes
Dicarboxylic Acids
Metabotropic Glutamate Receptors
Kainic Acid
Tetrodotoxin
Phorbol Esters
N-Methylaspartate
GABA-A Receptors

Keywords

  • Metabotropic receptors
  • Phorbol esters
  • Phosphoinositides
  • Trans-(±)-1-Amino-1,3-cyclopentane dicarboxylic acid
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Activation of p42 mitogen-activated protein kinase by glutamate receptor stimulation in rat primary cortical cultures. / Fiore, R. S.; Murphy, T. H.; Sanghera, J. S.; Pelech, S. L.; Baraban, Jay M.

In: Journal of Neurochemistry, Vol. 61, No. 5, 11.1993, p. 1626-1633.

Research output: Contribution to journalArticle

Fiore, R. S. ; Murphy, T. H. ; Sanghera, J. S. ; Pelech, S. L. ; Baraban, Jay M. / Activation of p42 mitogen-activated protein kinase by glutamate receptor stimulation in rat primary cortical cultures. In: Journal of Neurochemistry. 1993 ; Vol. 61, No. 5. pp. 1626-1633.
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