title: Brief low [Mg(2+)]o-induced Ca(2+) spikes inhibit subsequent prolonged exposure-induced excitotoxicity in cultured rat hippocampal neurons.
abstract: Reducing [Mg(2+)]o to 0.1 mM can evoke repetitive [Ca(2+)]i spikes and seizure activity, which induces neuronal cell death in a process called excitotoxicity. We examined the issue of whether cultured rat hippocampal neurons preconditioned by a brief exposure to 0.1 mM [Mg(2+)]o are rendered resistant to excitotoxicity induced by a subsequent prolonged exposure and whether Ca(2+) spikes are involved in this process. Preconditioning by an exposure to 0.1 mM [Mg(2+)]o for 5 min inhibited significantly subsequent 24 h exposure-induced cell death 24 h later (tolerance). Such tolerance was prevented by both the NMDA receptor antagonist D-AP5 and the L-type Ca(2+) channel antagonist nimodipine, which blocked 0.1 mM [Mg(2+)]o-induced [Ca(2+)]i spikes. The AMPA receptor antagonist NBQX significantly inhibited both the tolerance and the [Ca(2+)]i spikes. The intracellular Ca(2+) chelator BAPTA-AM significantly prevented the tolerance. The nonspecific PKC inhibitor staurosporin inhibited the tolerance without affecting the [Ca(2+)]i spikes. While Gö6976, a specific inhibitor of PKCα had no effect on the tolerance, both the PKCε translocation inhibitor and the PKCζ pseudosubstrate inhibitor significantly inhibited the tolerance without affecting the [Ca(2+)]i spikes. Furthermore, JAK-2 inhibitor AG490, MAPK kinase inhibitor PD98059, and CaMKII inhibitor KN-62 inhibited the tolerance, but PI-3 kinase inhibitor LY294,002 did not. The protein synthesis inhibitor cycloheximide significantly inhibited the tolerance. Collectively, these results suggest that low [Mg(2+)]o preconditioning induced excitotoxic tolerance was directly or indirectly mediated through the [Ca(2+)]i spike-induced activation of PKCε and PKCξ, JAK-2, MAPK kinase, CaMKII and the de novo synthesis of proteins.
authors.affiliation: Department of Physiology, College of Medicine, Dankook University, Cheonan 31116, Korea.
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.
authors.forename: Ji Seon
authors.initials: JS
authors.lastname: Yang
authors.affiliation: Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.; Catholic Neuroscience Institute, The Catholic University of Korea, Seoul 06591, Korea.
authors.forename: Shin Hee
authors.initials: SH
authors.lastname: Yoon
keywords: Ca2+ spikes; Excitotoxicity; Low [Mg2+]o preconditioning