Activation, involvement and nuclear translocation of c-Jun N-terminal protein kinase 1 and 2 in glutamate-induced apoptosis in cultured rat cortical neurons
Activation, involvement and nuclear translocation of c-Jun N-terminal protein kinase 1 and 2 in glutamate-induced apoptosis in cultured rat cortical neurons
复制标题
DOI：
10.1016/s0006-8993(02)03435-2
复制
发表时间：
2002-11-29
期刊：
影响因子：
2.9
通讯作者：
Zhang, GY
中科院分区：
文献类型：
--
作者：
Jiang, Q;Gu, ZL;Zhang, GY
先前的研究表明，c-Jun N-末端蛋白激酶1和2（JNK 1和2）在某些情况下的兴奋性毒性激活。在本研究中，JNK 1和2的激活，亚细胞分布，参与和上游调控谷氨酸诱导的兴奋性毒性在培养的大鼠皮层神经元进行了研究。如来自全细胞提取物的Western免疫印迹所示，虽然JNK 1和2没有显著改变，但活化的JNK 1和2在细胞内表达。（二磷酸化JNK 1和2，p-JNK 1和2）在暴露于50 μ M谷氨酸15分钟时迅速增加，并在暴露后12小时恢复到基础水平，随后在暴露后9-18小时通过DAPI（一种荧光DNA结合染料）染色检测到的凋亡样细胞死亡显著增加。用JNK 1和2反义寡核苷酸阻断p-JNK 1和2的增加可显著防止细胞死亡。阻断NMDA受体（谷氨酸受体的一种亚型）或蛋白激酶C（PKC）可在很大程度上阻止p-JNK 1和2的增加，并且每种阻断也在很大程度上阻止细胞死亡。联合阻断PKC和JNK 1和2对细胞死亡没有相加的保护作用。免疫细胞化学研究显示，在谷氨酸暴露15分钟时，整个细胞但主要是核的p-JNK 1和2的增加，连同轻度的血浆JNK 1和2的减少但核的大量增加，所有这些也在很大程度上被NMDA受体或PKC阻断所阻止。这些结果表明，主要下游的NMDA受体-PKC途径JNK 1和2被激活，核转位和因果关系参与谷氨酸诱导的兴奋性毒性，可能通过核升高的p-JNK 12。(C)2002 Elsevier Science B. V.保留所有权利。
Previous studies showed that c-Jun N-terminal protein kinase 1 and 2 (JNK1&2) were activated in some cases of excitotoxicity. In the present study, activation, subcellular distribution, involvement and upstream regulation of JNK1&2 were investigated in glutamate-induced excitotoxicity in cultured rat cortical neurons. As indicated by Western immunoblot from whole cellular extracts, while JNK1&2 were not significantly changed, the activated JNK1&2 (diphosphorylated JNK1&2, p-JNK1&2), were rapidly increased at 15 min exposure to 50 muM glutamate and reverted to basal level at 12 h after exposure, followed by a significant increase of apoptotic-like cell death as detected by DAPI (a fluorescent DNA binding dye) staining at 9-18 h after exposure. Blockage of the increase of p-JNK1&2 with JNKI&2 antisense oligodeoxynucleotides significantly prevented the cell death. The increase of p-JNK1&2 was largely prevented by blockage of NMDA receptor (a subtype of glutamate receptor) or protein kinase C (PKC), and each blockage also largely prevented the cell death. Combined blockage of PKC and JNK1&2 had no additive protective effect against cell death. Immunocytochemistry study showed at 15 min of glutamate exposure a whole cellular but mainly nuclear increase of p-JNK1&2, together with mild plasma decrease but large nuclear increase of JNK1&2, all of which were also largely prevented by blockage of NMDA receptor or PKC. These results suggested that mainly downstream of NMDA receptor-PKC pathway JNK1&2 were activated, nuclear translocated and causally involved in the glutamate-induced excitotoxicity, possibly through a nuclear elevation of p-JNK12. (C) 2002 Elsevier Science B.V. All rights reserved.