Elsevier

Neuroscience

Volume 76, Issue 2, January 1997, Pages 411-423
Neuroscience

Dual mode ofN-methyl-d-aspartate-induced neuronal death in hippocampal slice cultures in relation toN-methyl-d-aspartate receptor properties

https://doi.org/10.1016/S0306-4522(96)00403-4Get rights and content

Abstract

Hippocampal slices prepared from neonatal rats were cultured for several weeks, and excitotoxicity induced in CA1 pyramidal neurons byN-methyl-d-aspartate was evaluated at different culture stages. CA1 neurons cultured for one week exhibited cell death predominantly within 1–3 h after a 15-minN-methyl-d-aspartate insult (early death), whereas those cultured for three weeks showed cell death mainly a few hours to 24 h after the insult (delayed death). CA1 neurons cultured for two weeks were in a transitional state, expressing only weak early and delayed forms of cell death in response toN-methyl-d-aspartate. TheN-methyl-d-aspartate-induced early cell death observed in the one-week group depended on external Cl but did not require external Ca2+; rather, early cell death was enhanced in Ca2+-free solution. This early cell death was accompanied by cell swelling, but cell swelling alone produced by osmotic changes failed to induce cell death. There was no evidence that CA1 neurons in the one-week group were more responsive toN-methyl-d-aspartate than those in the two other groups. Delayed cell death examined in the three-week group depended on external Ca2+ but was independent of Cl. The density ofN-methyl-d-aspartate-induced whole-cell currents recorded from CA1 neurons in Mg2+-free solution remained unchanged during three weeks of culture. However, theN-methyl-d-aspartate receptor channel in the three-week group was more resistant to Mg2+ block than that in the one- or two-week group. The incidence ofN-methyl-d-aspartate-induced delayed cell death was higher in the three-week group than in the two-week group in normal solution but not in Mg2+-free solution. Thus, Mg2+, block-resistant properties of theN-methyl-d-aspartate receptor channel acquired during prolonged culture may account for the high incidence ofN-methyl-d-aspartate-induced delayed cell death in the three-week group. However, theN-methyl-d-aspartate receptor subunits expressed in the CA1 subfield did not show any feature specific to the three-week group.

These results show that two mechanistically distinct modes ofN-methyl-d-aspartate-induced neuronal death are manifested differentially at different culture stages, depending on the intrinsic neuron properties (i.e. early cell death) and on the properties or the responsiveness of theN-methyl-d-aspartate receptor (i.e. delayed cell death).

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