Excitotoxicity in vitro by NR2A- and NR2B-containing NMDA receptors.
Excitotoxicity, exacerbating acute brain damage from brain trauma or stroke, is mediated in part by excessive Ca(2+)-influx from prolonged NMDA receptor activation. However, the contribution to excitotoxicity by each of the main NMDAR subtypes in glutamatergic forebrain neurons, the NR2A- and NR2B-types, has remained enigmatic. Here, we investigated this issue by use of pharmacological and genetic tools in cultured cortical neurons. In wild-type neurons the contribution of the NMDA receptor subtypes to excitotoxicity changed with the age of the cultures. The blockade of NR2B-containing NMDA receptors prevented NMDA-mediated toxicity in young cultures after 14days in vitro (DIV14), but both subtypes triggered excitotoxicity in older (DIV21) cultures. Notably, blocking either of the two subtypes failed to prevent NMDA-elicited cell death, indicating that the remaining subtype triggers cell demise. Intriguingly, a neuroprotective aspect of the NR2A subtype became apparent at submaximal NMDA concentration only at DIV21. The NR2A subtype mediated NMDA toxicity as well as partial protection only if it carried a functional C-terminal domain. Upon deletion of this domain in the NR2A subtype, excitotoxicity was mediated entirely via the NR2B subtype, both at DIV14 and DIV21. Our findings predict that successful therapeutic intervention in stroke based on currently available NMDA receptor subtype-selective blockers is unlikely.
von Engelhardt
Jakob
J
IZN and Department of Clinical Neurobiology, University Hospital of Neurology, INF 364, 69120 Heidelberg, Germany.
Coserea
Irinel
I
Pawlak
Verena
V
Fuchs
Elke C
EC
Köhr
Georg
G
Seeburg
Peter H
PH
Monyer
Hannah
H
D000704:Analysis of Variance; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004622:Embryo, Mammalian; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000502:physiology; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010880:Piperidines / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D016194:Receptors, N-Methyl-D-Aspartate / Q000172:deficiency / Q000502:physiology*; D013997:Time Factors
C498554:5-(alpha-methyl-4-bromobenzylamino)phosphonomethyl-1,4-dihydroquinoxaline-2,3-dione; D002121:Calcium Channel Blockers; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; C120997:NR2A NMDA receptor; C121001:NR2B NMDA receptor; D010880:Piperidines; D011810:Quinoxalines; D016194:Receptors, N-Methyl-D-Aspartate; D016202:N-Methylaspartate; C010739:ifenprodil