Activation of a caspase 3-related cysteine protease is required for glutamate-mediated apoptosis of cultured cerebellar granule neurons
Activation of a caspase 3-related cysteine protease is required for glutamate-mediated apoptosis of cultured cerebellar granule neurons
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DOI：
10.1073/pnas.94.21.11657
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发表时间：
1997-10-14
影响因子：
11.1
通讯作者：
Paul, SM
中科院分区：
文献类型：
--
作者：
Du, YS;Bales, KR;Paul, SM
过度刺激 N-甲基-D-天冬氨酸 (NMDA) 受体引起的神经毒性部分是由于细胞内 Ca2+ 持续升高；然而，对于最终导致细胞死亡的细胞内事件知之甚少。在这里，我们表明，相对较低浓度的谷氨酸过度刺激 NMDA 受体会诱导培养的小脑颗粒神经元 (CGN) 凋亡，并且 CGN 不需要新的 RNA 或蛋白质合成。然而，谷氨酸诱导的 CGN 凋亡与浓度和时间相关。 白细胞介素 1 β 转换酶 (ICE)/CED-3 相关蛋白酶、CPP32/Yama/apopain（现称为 caspase 3）的依赖性激活，此外，CGN 暴露于谷氨酸后 caspase 3 激活的时间过程与细胞凋亡的发展平行，此外，谷氨酸诱导的 CGN 细胞凋亡几乎完全被选择性细胞阻断 caspase 3 的渗透性四肽抑制剂 Ac-DEVD-CHO，但不是 ICE (caspase 1) 抑制剂 Ac-YVAD-CHO，来自谷氨酸暴露的 CGN 的胞质提取物的蛋白质印迹揭示了 caspase 3 底物聚（ADP-核糖）聚合酶的裂解，以及 caspase 3 前体蛋白水解加工成活性亚基，我们的 数据表明，谷氨酸诱导的 CGN 细胞凋亡是由 ICF/CED-3 相关半胱氨酸蛋白酶 caspase 3 的翻译后激活介导的。
Neurotoxicity induced by overstimulation of N-methyl-D-aspartate (NMDA) receptors is due, in part, to a sustained rise in intracellular Ca2+; however, little is known about the ensuing intracellular events that ultimately result in cell death, Here we show that overstimulation of NMDA receptors by relatively low concentrations of glutamate induces apoptosis of cultured cerebellar granule neurons (CGNs) and that CGNs do not require new RNA or protein synthesis, Glutamate-induced apoptosis of CGNs is, however, associated with a concentration-and time dependent activation of the interleukin 1 beta-converting enzyme (ICE)/CED-3-related protease, CPP32/Yama/apopain (now designated caspase 3), Further, the time course of caspase 3 activation after glutamate exposure of CGNs parallels the development of apoptosis, Moreover, glutamate-induced apoptosis of CGNs is almost completely blocked by the selective cell permeable tetrapeptide inhibitor of caspase 3, Ac-DEVD-CHO but not by the ICE (caspase 1) inhibitor, Ac-YVAD-CHO, Western blots of cytosolic extracts from glutamate-exposed CGNs reveal both cleavage of the caspase 3 substrate, poly(ADP-ribose) polymerase, as well as proteolytic processing of pro-caspase 3 to active subunits, Our data demonstrate that glutamate-induced apoptosis of CGNs is mediated by a posttranslational activation of the ICF/CED-3-related cysteine protease caspase 3.