Mitochondrial transmembrane potential and free radical production in excitotoxic neurodegeneration

被引:47
作者
Prehn, JHM [1 ]
机构
[1] Univ Marburg, Inst Pharmakol & Toxikol, FB 16, D-35032 Marburg, Germany
关键词
rat hippocampal neurons; glutamate; hydroethidine; rhodamine-123; superoxide; cell death; digital videomicroscopy;
D O I
10.1007/PL00005173
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Excitotoxic cell death is involved in many forms of acute and chronic neurodegeneration. We induced excitotoxic cell death in cultured rat hippocampal neurons by brief exposure to two selective glutamate receptor agonists with different neurotoxic potencies, N-methyl-D-aspartate (NMDA) and kainate (KA). Digital video imaging was performed during exposure to the agonists to monitor free radical production and changes in mitochondrial transmembrane potential, Psi(m). Brief exposure to NMDA (10 min) induced significant cell death in the hippocampal neurons reaching a maximum at a concentration of 300 mu M (57.2+/-2.6% cell death; P<0.001). In parallel imaging experiments we found that exposure to NMDA (300 mu M, 10 min) induced a significant increase in superoxide production monitored with the oxidation-sensitive probe, hydroethidine (increase of 280+/-33% above baseline, P<0.001). Rhodamine-123-based imaging revealed a loss of Psi(m) in 70.1+/-10.1% of the hippocampal neurons during the exposure to NMDA. In contrast to NMDA, brief exposure to KA (10 min) produced limited neurotoxicity reaching a maximum at a concentration of 100 mu M (10.2+/-4.0% cell death: P<0.05. Exposure to KA (100 mu M, 10 min) also caused a significant increase in superoxide production. This increase, however, was significantly less pronounced than that produced by NMDA (increase of 94+/-17% above baseline: P<0.001 compared to controls or NMDA-exposed cultures). Moreover, rhodamine-123-based imaging revealed that KA (100 mu M) caused a collapse of Psi(m) in only 13.5+/-1.4% of the hippocampal neurons. In conclusion, the present study demonstrates that early changes in intracellular superoxide production and Psi(m) relate to neuronal survival outcome in excitotoxic cell death.
引用
收藏
页码:316 / 322
页数:7
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