遗传性癫痫大鼠海马内GAT-1和GAT-3表达上调  

Up-regulation of GAT-1,GAT-3 in genetic epileptic rat hippocampus

作　　者：毛小元[1] 俞军龄[1] 闵冬雨[1] 郭凤[1] 刘睿[1] 吴坤灿[1] 张宇轩[1] 谢妮[1] 蔡际群[1] 

机构地区：[1]中国医科大学药学院药物毒理学教研室,辽宁沈阳110001

出　　处：《中国药理学通报》2010年第5期629-633,共5页Chinese Pharmacological Bulletin

基　　金：教育部高校博士学科专项科研基金资助项目(No20060159003)

摘　　要：目的探讨遗传性癫痫大鼠(TRM)海马内GAT-1和GAT-3mRNA与蛋白的表达。方法应用RT-PCR技术检测GAT-1和GAT-3mRNA的表达;应用Westernblot方法检测GAT-1和GAT-3蛋白的表达;应用免疫组化方法定位GAT-1和GAT-3蛋白的分布。结果TRM大鼠海马内GAT-1mR-NA和蛋白表达均高于正常Wistar大鼠(P<0.01);GAT-3mRNA表达高于正常Wistar大鼠(P<0.05);GAT-3蛋白表达亦高于正常Wistar大鼠(P<0.01);免疫组化结果显示GAT-1在TRM和Wistar大鼠海马的CA1、CA3和DG区均有明显分布,且在TRM鼠海马的CA1、CA3和DG区中均比在正常Wistar鼠中明显;而GAT-3蛋白只在TRM和Wistar大鼠海马的CA1区中表达丰富,同时,和正常Wistar鼠比较,TRM鼠海马CA1区中GAT-3蛋白的表达明显增加。结论TRM大鼠海马内GAT-1和GAT-3表达上调可能促进了癫痫的发生,可能是癫痫治疗的靶点。Aim To investigate mRNA and protein expression of GAT-1 and GAT-3 in hippocampus of tremor rats（TRMs）and Wistar rats as the control groups.Methods Total RNA was extracted from the whole hippocampus tissues of TRMs and Wistar rats and mRNA expressions of GAT-1,GAT-3 were analysed byRT-PCR;the expressions of GAT-1 and GAT-3 at the protein level were detected by Western blot and immunohistochemistry.Results The expressions of GAT-1 at mRNA and protein levels were both significantly increased compared with the control group（P〈0.01）;we also observed the marked increase of GAT-3 mRNA and protein in the hippocampus of TRM（P〈0.05 and P〈0.01,respectively）.It was obviously observed that GAT-1 protein in TRMs and Wistar rats were both localized widely in hippocampus by immunohistochemistry including CA1,CA3 and DG regions.However,in terms of GAT-3,a weak immunoreaction appeared in CA3 and DG regions except for the evident observation in the CA1 region of hippocampus.Conclusions The up-regulation of GAT-1 and GAT-3 in TRM hippocampus may contribute to the generation of seizure which is likely to provide the potential therapeutic targets for genetic epilepsy.

关 键 词：GAT-1 GAT-3 TRM大鼠 癫痫 RT-PCR 免疫组化 WESTERN BLOT 

分 类 号：R-332[医药卫生] R322.81