Excitotoxic superoxide production and neuronal death require both ionotropic and non-ionotropic NMDA receptor signaling

Angela M. Minnella, Jerry X. Zhao, Xiangning Jiang, Emil Jakobsen, Fuxin Lu, Long Wu, Jamel El-Benna, John Gray, Raymond A. Swanson

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2 Scopus citations

Abstract

NMDA-type glutamate receptors (NMDAR) trigger superoxide production by neuronal NADPH oxidase-2 (NOX2), which if sustained leads to cell death. This process involves Ca 2+ influx through NMDAR channels. By contrast, comparable Ca 2+ influx by other routes does not induce NOX2 activation or cell death. This contrast has been attributed to site-specific effects of Ca 2+ flux through NMDAR. Here we show instead that it stems from non-ionotropic signaling by NMDAR GluN2B subunits. To evaluate non-ionotropic effects, mouse cortical neurons were treated with NMDA together with 7-chlorokynurenate, L-689,560, or MK-801, which block Ca 2+ influx through NMDAR channels but not NMDA binding. NMDA-induced superoxide formation was prevented by the channel blockers, restored by concurrent Ca 2+ influx through ionomycin or voltage-gated calcium channels, and not induced by the Ca 2+ influx in the absence of NMDAR ligand binding. Neurons expressing either GluN2B subunits or chimeric GluN2A/GluN2B C-terminus subunits exhibited NMDA-induced superoxide production, whereas neurons expressing chimeric GluN2B/GluN2A C-terminus subunits did not. Neuronal NOX2 activation requires phosphoinositide 3-kinase (PI3K), and NMDA binding to NMDAR increased PI3K association with NMDA GluN2B subunits independent of Ca 2+ influx. These findings identify a non-ionotropic signaling pathway that links NMDAR to NOX2 activation through the C-terminus domain of GluN2B.

Original languageEnglish (US)
Article number17522
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

ASJC Scopus subject areas

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    Minnella, A. M., Zhao, J. X., Jiang, X., Jakobsen, E., Lu, F., Wu, L., El-Benna, J., Gray, J., & Swanson, R. A. (2018). Excitotoxic superoxide production and neuronal death require both ionotropic and non-ionotropic NMDA receptor signaling. Scientific Reports, 8(1), [17522]. https://doi.org/10.1038/s41598-018-35725-5