Up-regulated ephrinB3/EphB3 expression in intractable temporal lobe epilepsy patients and pilocarpine induced experimental epilepsy rat model
Up-regulated ephrinB3/EphB3 expression in intractable temporal lobe epilepsy patients and pilocarpine induced experimental epilepsy rat model
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DOI：
10.1016/j.brainres.2016.02.035
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发表时间：
2016-05-15
期刊：
影响因子：
2.9
通讯作者：
Chen, Yangmei
中科院分区：
文献类型：
--
作者：
Huang, Hao;Li, Ruohan;Chen, Yangmei
EphB家族受体酪氨酸激酶与细胞表面结合的ephrinB配体合作，在中枢神经系统（CNS）树突棘形态发生、轴突引导、突触发生、突触重组和可塑性的维持中发挥关键作用。然而，ephrinB/EphB在难治性颞叶癫痫（TLE）中的表达模式和癫痫发生过程中潜在的分子机制尚不清楚。本研究采用实时定量聚合酶链反应（RT-qPCR）、免疫组织化学、双标记免疫荧光和Western blot方法研究ephrinB/EphB在顽固性TLE患者和氯化锂-匹罗卡品诱导的TLE大鼠中的表达模式和细胞分布。与对照组相比，顽固性TLE患者和TLE大鼠中ephrinB3和EphB3 mRNA表达量显著上调，而顽固性TLE患者和TLE大鼠中ephrinB1/2和EphB1/2/4/6 mRNA表达趋势不一致。Western blot分析和半定量免疫组织化学证实，顽固性TLE患者和TLE大鼠中ephrinB3和EphB3蛋白水平上调。同时，双标记免疫荧光显示ephrinB3主要表达于胶质细胞和神经元的胞质和突起，EphB3主要表达于神经元的胞质。综上所述，ephrinB3/EphB3在难治性TLE患者和实验性TLE大鼠中表达上调，提示ephrinB3/EphB3可能参与了TLE的发病机制。(C) 2016 Elsevier B.V.版权所有
EphB family receptor tyrosine kinases, in cooperation with cell surface-bound ephrinB ligands, play a critical role in maintenance of dendritic spine morphogenesis, axons guidance, synaptogenesis, synaptic reorganization and plasticity in the central nervous system (CNS). However, the expression pattern of ephrinB/EphB in intractable temporal lobe epilepsy (TLE) and the underlying molecular mechanisms during epileptogenesis remain poorly understood. Here we investigated the expression pattern and cellular distribution of ephrinB/EphB in intractable TLE patients and lithium chloride-pilocarpine induced TLE rats using real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry, double-labeled immunofluorescence and Western blot analysis. Compared to control groups, ephrinB3 and EphB3 mRNA expression were significantly up regulated in intractable TLE patients and TLE rats, while the mRNA expression trend of ephrinB1/2 and EphB1/2/4/6 in intractable TLE patients and TLE rats were inconsistent. Western blot analysis and semi-quantitative immunohistochemistry confirmed that ephrinB3 and EphB3 protein level were up-regulated in intractable TLE patients and TLE rats. At the same time, double-labeled immunofluorescence indicate that ephrinB3 was expressed mainly in the cytoplasm and protrusions of glia and neurons, while EphB3 was expressed mainly in the cytoplasm of neurons. Taken together, up-regulated expression of ephrinB3/EphB3 in intractable TLE patients and experimental TLE rats suggested that ephrinB3/EphB3 might be involved in the pathogenesis of TLE. (C) 2016 Elsevier B.V. All rights reserved.