Expression of death-associated protein kinase and recruitment to the tumor necrosis factor signaling pathway following brief seizures
Expression of death-associated protein kinase and recruitment to the tumor necrosis factor signaling pathway following brief seizures
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DOI：
10.1046/j.1471-4159.2003.01934.x
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发表时间：
2003-09-01
影响因子：
4.7
通讯作者：
Simon, RP
中科院分区：
文献类型：
--
作者：
Henshall, DC;Araki, T;Simon, RP
死亡相关蛋白（DAP）激酶是钙调节的，已知在死亡受体下游发挥作用，促使我们研究其在脑缺血诱导的神经元死亡机制中的作用。短暂的癫痫发作局灶性诱发大鼠，引起海马CA 3子区内的神经元死亡，并在较小程度上，皮质。蛋白质印迹法证实了DAP激酶在海马和皮质中的表达，预测的重量类似于160 kDa。免疫组化显示癫痫发作触发了CA 3和皮质内DAP激酶表达细胞数量的显着增加，而不影响抗癫痫CA 2或齿状回内的细胞计数。DAP激酶表达细胞的数量增加与特定模式的损伤引起的癫痫发作活动，电图定义。癫痫发作引起早期增加DAP激酶结合肌动蛋白，并与钙调蛋白。免疫共沉淀研究还显示癫痫发作触发DAP激酶与肿瘤坏死因子受体1和Fas相关死亡结构域蛋白的结合，与caspase-8蛋白水解相当。相反，在海马的存活区域内，DAP激酶与分子伴侣14-3-3相互作用。这些数据表明，DAP激酶参与了脑缺血诱导的神经元死亡过程中激活的分子通路。
Death-associated protein (DAP) kinase is calcium-regulated and known to function downstream of death receptors, prompting us to examine its role in the mechanism of seizure-induced neuronal death. Brief seizures were focally evoked in rats, eliciting neuronal death within the CA3 subfield of the hippocampus, and to a lesser extent, cortex. Western blotting confirmed expression of DAP kinase within hippocampus and cortex at the predicted weight of similar to160 kDa. Immunohistochemistry revealed seizures triggered a significant increase in numbers of DAP kinase-expressing cells within CA3 and cortex, without affecting cell counts within seizure-resistant CA2 or the dentate gyrus. Numbers of DAP kinase-expressing cells were increased in relation to specific patterns of injury-causing seizure activity, electrographically defined. Seizures caused an early increase in DAP kinase binding to actin, and association with calmodulin. Co-immunoprecipitation studies also revealed seizures triggered binding of DAP kinase to the tumor necrosis factor receptor 1 and the Fas-associated death domain protein, commensurate with caspase-8 proteolysis. In contrast, within surviving fields of the hippocampus, DAP kinase interacted with the molecular chaperone 14-3-3. These data suggest DAP kinase is involved in the molecular pathways activated during seizure-induced neuronal death.