Wu Liwen, Yin Fei, Peng Jing, He Fang, Zhang Ciliu, Deng Xiaolu, Wang Guoli
Department of Pediatrics, Xiangya Hospital, Central South University, Changsha, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2013 Jun;38(6):560-9. doi: 10.3969/j.issn.1672-7347.2013.06.003.
To examine the changes of morphology and differentially expressed proteins in hippocampus at the latent stage of chronic mesial temporal lobe epilepsy (MTLE) in immature rats, and to explore the global mechanism of chronic MTLE at a new point.
MTLE models of immature rats were induced by lithium-pilocarpine. The rats were divided into 2 groups randomly: a control group (n=20) and an MTLE model group (n=20). At the latent stage, nissl and Timm staining were performed to evaluate the cell loss and mossy fiber sprouting. The differentially expressed proteins were separated by 2-dimensional polyacrylamide gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF-MS) technology. Western blot was used to determine the differentially expression levels of partial proteins.
Neuron loss and abnormal mossy fiber sprouting were obviously observed in the hippocampus in the MTLE model group; 2-DE patterns of hippocampus of the MTLE model group in latent stage and the control group were established. Thirty-one differential proteins were identified by MALDI-TOF-MS, which were categorized into several groups by biological functions: synaptic and neurotransmitter release related proteins, cytoskeletal proteins, cell junctions proteins, energy metabolism and mitochondrial proteins, biological enzymes, cellular structure related proteins, signal regulating molecular and others. The expression levels of partial proteins determined by Western blot were similar to the changes of proteomics.
The differentially expressed proteins of synapse-related proteins such as dynamin-1, neurogranin and ubiquitin, which cause the synapse reorganization and mossy fiber terminal sprouting related to the formation of abnormal excitatory network, probably play critic roles in the mechanism of MTLE.
观察幼龄大鼠慢性内侧颞叶癫痫(MTLE)潜伏期海马形态及差异表达蛋白质的变化,从新的角度探讨慢性MTLE的整体机制。
采用锂-匹罗卡品诱导幼龄大鼠MTLE模型。将大鼠随机分为2组:对照组(n=20)和MTLE模型组(n=20)。在潜伏期,进行尼氏染色和Timm染色以评估细胞丢失和苔藓纤维发芽。采用二维聚丙烯酰胺凝胶电泳(2-DE)结合基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)技术分离差异表达蛋白质。用蛋白质免疫印迹法检测部分蛋白质的差异表达水平。
MTLE模型组海马明显可见神经元丢失和异常苔藓纤维发芽;建立了MTLE模型组潜伏期海马与对照组海马的2-DE图谱。通过MALDI-TOF-MS鉴定出31种差异蛋白质,按生物学功能分为几类:突触和神经递质释放相关蛋白、细胞骨架蛋白、细胞连接蛋白、能量代谢和线粒体蛋白、生物酶、细胞结构相关蛋白、信号调节分子等。蛋白质免疫印迹法检测的部分蛋白质表达水平变化与蛋白质组学变化相似。
发动蛋白-1、神经颗粒蛋白和泛素等与突触相关的差异表达蛋白质导致突触重组和苔藓纤维终末发芽,与异常兴奋性网络形成有关,可能在MTLE机制中起关键作用。