GLUTAMATE INDUCES THE PRODUCTION OF REACTIVE OXYGEN SPECIES IN CULTURED FOREBRAIN NEURONS FOLLOWING NMDA RECEPTOR ACTIVATION
GLUTAMATE INDUCES THE PRODUCTION OF REACTIVE OXYGEN SPECIES IN CULTURED FOREBRAIN NEURONS FOLLOWING NMDA RECEPTOR ACTIVATION
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DOI：
10.1523/jneurosci.15-05-03318.1995
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发表时间：
1995-05-01
影响因子：
5.3
通讯作者：
HASTINGS, TG
中科院分区：
文献类型：
--
作者：
REYNOLDS, IJ;HASTINGS, TG
用荧光染料2‘，7’-二氯二氢荧光素(DCF-H-2)研究了谷氨酸在培养的胎鼠前脑神经元产生活性氧(ROS)中的作用。加入兴奋性毒性浓度的谷氨酸(100mM)后，细胞DCF的荧光普遍下降，并伴有细胞体边缘局部区域的荧光增强，这种增强可在谷氨酸作用2-4分钟内观察到，荧光的增强依赖于NMDA受体的激活和钙离子的进入，并被线粒体质子离子载体氰基乙酰对三氟甲氧基苯肼(FCCP)所阻断。更多的研究表明，荧光的普遍下降是由于细胞内的酸化。这些研究表明，线粒体在谷氨酸兴奋性毒性相关的ROS的产生中起着关键作用，并进一步证明了在单细胞水平上测量ROS产生的可行性。
Using the fluorescent dye 2',7'-dichlorodihydrofluorescein (DCF-H-2) we investigated the role of glutamate in the production of reactive oxygen species (ROS) in cultured neurons from fetal rat forebrain. The addition of an excitotoxic concentration of glutamate (100 mu M) produced a generalized decrease in cellular DCF fluorescence accompanied by local areas of increased fluorescence around the margins of the cell body that could be observed within 2-4 min of glutamate exposure, Increases in fluorescence were dependent on NMDA receptor activation and Ca2+ entry and were blocked by the mitochondrial proton ionophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP). Additional studies suggested that the generalized decrease in fluorescence was due to intracellular acidification. These studies suggest a critical role for mitochondria in the production of ROS in association with glutamate excitotoxicity, and additionally demonstrate the feasibility of measuring the production of ROS at the level of the single cell.