Glutamate receptor-mediated currents and toxicity in embryonal carcinoma cells.
Glutamate receptor-mediated currents and toxicity in embryonal carcinoma cells.
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胚胎癌细胞中谷氨酸受体介导的电流和毒性。
DOI：
10.1002/neu.480240904
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Choi,DW
中科院分区：
文献类型：
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作者：
Turetsky,DM;Huettner,JE;Gottlieb,DI;Goldberg,MP;Choi,DW
虽然原代神经元细胞培养物已用于研究兴奋毒性，但仍需要开发表现出谷氨酸受体介导的死亡的细胞系。 P19 小鼠胚胎癌细胞暴露于视黄酸并铺板到培养的小鼠皮质神经胶质细胞层上，分化成对神经丝和神经元特异性烯醇化酶具有免疫反应性的神经元样元件。全细胞记录显示，细胞外应用 NMDA 或红藻氨酸会产生内向电流。 NMDA 诱导的电流表现出镁的电压依赖性阻断，需要甘氨酸才能最大激活，并被 NMDA 拮抗剂地佐西平阻断。红藻氨酸诱导的电流被 AMPA/红藻氨酸受体拮抗剂 CNQX 阻断。暴露于 500 μMNMDA 24 小时可破坏大多数 P19 细胞（EC50 约 70 μM）；地佐西平或 D-APV 预防了死亡。暴露于 500 μMkainate 也导致 CNQX 减少了广泛的死亡。因此，分化的 P19 细胞表现出兴奋性氨基酸反应和对兴奋性毒性的脆弱性，这是中枢神经系统神经元的特征。这些细胞可能提供一个遗传开放系统，可用于在分子水平上研究谷氨酸受体介导的现象。 © 1993 约翰威利父子公司。
While primary neuronal cell cultures have been used to investigate excitotoxicity, development of cell lines exhibiting glutamate receptor‐mediated death is desirable. P19 mouse embryonal carcinoma cells, exposed to retinoic acid and plated onto a layer of cultured mouse cortical glial cells, differentiated into neuron‐like elements immunoreactive for neurofilaments and neuron‐specific enolase. Whole‐cell recordings revealed inward currents in response to extracellular application of either NMDA or kainate. The NMDA‐induced currents exhibited a voltage‐dependent blockade by magnesium, required glycine for maximal activation, and were blocked by the NMDA antagonist dizocilpine. Kainate‐induced currents were blocked by the AMPA/kainate receptor antagonist CNQX. Exposure to 500 μMNMDA for 24 h destroyed most P19 cells (EC50approximately 70 μM); death was prevented by dizocilpine or D‐APV. Exposure to 500 μMkainate also resulted in widespread death reduced by CNQX. Thus differentiated P19 cells exhibited both excitatory amino acid responses and vulnerability to excitotoxicity, characteristic of CNS neurons. These cells may provide a genetically open system useful for studying glutamate receptor‐mediated phenomena at a molecular level. © 1993 John Wiley & Sons, Inc.