Rapid phosphorylation of histone H2A.X following ionotropic glutamate receptor activation

Samantha L. Crowe, Vilen A. Movsesyan, Timothy J. Jorgensen, Alexei Kondratyev

Research output: Contribution to journalArticle

Abstract

Excessive activation of ionotropic glutamate receptors increases oxidative stress, contributing to the neuronal death observed following neurological insults such as ischemia and seizures. Post-translational histone modifications may be key mediators in the detection and repair of damage resulting from oxidative stress, including DNA damage, and may thus affect neuronal survival in the aftermath of insults characterized by excessive glutamate release. In non-neuronal cells, phosphorylation of histone variant H2A.X (termed γ-H2AX) occurs rapidly following DNA double-strand breaks. We investigated γ-H2AX formation in rat cortical neurons (days in vitro 14) following activation of N-methyl-d-aspartate (NMDA) or α-amino-3-hydroxyl-5-methyl- 4-isoxazolepropionic acid (AMPA)/kainate glutamate receptors using fluorescent immunohistochemical techniques. Moreover, we evaluated the co-localization of γ-H2AX 'foci' with Mre11, a double-strand break repair protein, to provide further evidence for the activation of this DNA damage response pathway. Here we show that minimally cytotoxic stimulation of ionotropic glutamate receptors was sufficient to evoke γ-H2AX in neurons, and that NMDA-induced γ-H2AX foci formation was attenuated by pretreatment with the antioxidant, Vitamin E, and the intracellular calcium chelator, BAPTA-AM. Moreover, a subset of γ-H2AX foci co-localized with Mre11, indicating that at least a portion of γ-H2AX foci is damage dependent. The extent of γ-H2AX induction following glutamate receptor activation corresponded to the increases we observed following conventional DNA damaging agents [i.e. non-lethal doses of gamma-radiation (1 Gy) and hydrogen peroxide (10 μm)]. These data suggest that insults not necessarily resulting in neuronal death induce the DNA damage-evoked chromatin modification, γ-H2AX, and implicate a role for histone alterations in determining neuronal vulnerability following neurological insults.

Original languageEnglish (US)
Pages (from-to)2351-2361
Number of pages11
JournalEuropean Journal of Neuroscience
Volume23
Issue number9
DOIs
StatePublished - May 2006
Externally publishedYes

Fingerprint

Ionotropic Glutamate Receptors
Histones
DNA Damage
Phosphorylation
Glutamate Receptors
Aspartic Acid
Oxidative Stress
Histone Code
Kainic Acid Receptors
Neurons
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
Double-Stranded DNA Breaks
Gamma Rays
Post Translational Protein Processing
Vitamin E
Hydroxyl Radical
Hydrogen Peroxide
Chromatin
Glutamic Acid
Seizures

Keywords

  • γ-H2AX
  • Cortical neurons
  • Glutamate receptors
  • Histone modification
  • Rat

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Rapid phosphorylation of histone H2A.X following ionotropic glutamate receptor activation. / Crowe, Samantha L.; Movsesyan, Vilen A.; Jorgensen, Timothy J.; Kondratyev, Alexei.

In: European Journal of Neuroscience, Vol. 23, No. 9, 05.2006, p. 2351-2361.

Research output: Contribution to journalArticle

Crowe, Samantha L. ; Movsesyan, Vilen A. ; Jorgensen, Timothy J. ; Kondratyev, Alexei. / Rapid phosphorylation of histone H2A.X following ionotropic glutamate receptor activation. In: European Journal of Neuroscience. 2006 ; Vol. 23, No. 9. pp. 2351-2361.
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