NMDA-induced neurodegeneration in mouse hippocampal slice cultures. After 6 days in culture, hippocampal slice cultures were treated with concentrations of 0 (control), 10, 15, 25 and 50 μM NMDA. Treatment with NMDA clearly induced cell death in the slice cultures as determined by propidium iodide uptake (PI; red), which co-localized with the neuronal nuclear marker NeuN (C, white arrow), indicating that NMDA specifically induced neuronal cell death. A concentration-dependent vulnerability towards NMDA was observed as neurons of the CA1 region were most sensitive to the NMDA-treatment (with 67.1% cell death at 10 μM NMDA), followed by the CA3 (41.7% at 15 μM NMDA) and finally the DG, which showed relatively low levels of cell death even at 50 μM NMDA (30.7%). Control slice cultures showed hardly any cell death (< 1%, B). Treatment of slice cultures with the NMDA-antagonist MK-801 (30 μM) for 1 hour prior to NMDA-treatment completely blocked NMDA-induced neuronal cell death. The percentages of neuronal cell death per neuronal cell layer (DG/CA3/CA1) were quantified and are represented in figure 5. Scale bars indicate 100 μm (A) and 25 μm (B,C).
Vinet et al. Journal of Neuroinflammation 2012 9:27 doi:10.1186/1742-2094-9-27