Apoptosis and the receptor specificity of its mechanisms during the neurotoxic action of glutamate.
Apoptosis and the receptor specificity of its mechanisms during the neurotoxic action of glutamate.
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DOI：
10.1007/s11055-009-9141-7
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发表时间：
2009-05-01
影响因子：
--
通讯作者：
Antonov, S M
中科院分区：
文献类型：
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作者：
Evstratova, A A;Mironova, E V;Antonov, S M
研究了谷氨酸和谷氨酸受体选择性激动剂N-甲基-D-天冬氨酸(NMDA)和红藻氨酸诱导的神经变性过程中坏死与凋亡的比例以及凋亡的机制。实验在大鼠大脑皮层神经元的原代培养物（体外七天）上进行。使用活体荧光快速测试并用吖啶橙和溴化乙锭染色来鉴定细胞凋亡和坏死。免疫细胞化学与共聚焦显微镜相结合用于观察凋亡蛋白。激动剂（240 分钟）通过这两个过程引起神经元死亡，但 NMDA 和红藻氨酸诱导神经变性时坏死细胞的比例显着低于谷氨酸诱导神经变性时。 3 mM 谷氨酸的神经毒性作用是通过 α-氨基-3-(3-羟基-5-甲基异恶唑-4-基)丙酸酯 (AMPA) 和红藻氨酸受体介导的，因为它被 6-氰基-7-硝基喹喔啉-2,3-二酮 (CNQX) 阻断。由于细胞凋亡诱导因子 (AIF) 对神经元细胞核的直接作用，NMDA 受体的激活导致细胞凋亡的发生，而无需半胱天冬酶的参与。 AMPA-红藻氨酸受体的激活伴随着通过半胱天冬酶依赖性途径发生的细胞凋亡。因此，这些数据确定了谷氨酸神经毒性作用期间细胞凋亡机制的受体依赖性。
The ratio of necrosis to apoptosis and the mechanisms of apoptosis were studied during neurodegeneration induced by glutamate and selective agonists of glutamate receptors - N-methyl-D-aspartate (NMDA) and kainate. Experiments were performed on primary cultures (seven days in vitro) of rat cerebral cortex neurons. Apoptosis and necrosis were identified using a vital fluorescence rapid test with staining with acridine orange and ethidium bromide. Immunocytochemistry in combination with confocal microscopy was used to visualize apoptotic proteins. Agonists (240 min) caused neuron death via both processes, though the proportion of necrotic cells when neurodegeneration was induced by NMDA and kainate was significantly less than when neurodegeneration was induced with glutamate. The neurotoxic effect of 3 mM glutamate was mediated via alpha-amino-3-(3-hydroxy-5-methylisoxazole-4-yl)propionate (AMPA) and kainate receptors, as it was blocked by 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX). Activation of NMDA receptors led to the development of apoptosis without involvement of caspases, due to the direct action of apoptosis-inducing factor (AIF) on neuron nuclei. Activation of AMPA-kainate receptors was accompanied by the development of apoptosis via the caspase-dependent pathway. Thus, these data identified the receptor dependence of the mechanisms of apoptosis during the neurotoxic action of glutamate.