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Cx43基因干扰对大鼠星形胶质细胞谷氨酸基础释放的影响
王凌云，田衍平,牛建钦，肖 岚（400038，重庆，第三军医大学基础部组织学与胚胎学教研室，重庆市神经科学重点实验室）
[摘要] 目的 构建能有效抑制大鼠星形胶质细胞连接蛋白43（connexin 43，Cx43）基因表达的siRNA载体，并检测抑制Cx43表达对星形胶质细胞谷氨酸释放的影响。方法 合成针对大鼠Cx43基因的siRNA寡核苷酸，。质粒通过电转方法转染原代星形胶质细胞 (astrocyte, AST）后，用免疫荧光染色及Western blot检测转染干扰质粒对大鼠ASTs Cx43蛋白表达的影响。用谷氨酸ELISA定量试剂盒检测释放至培养基中谷氨酸的浓度,并用DAPI染色进行细胞计数。结果 测序结果显示，Cx43基因的目的片段正确插入载体，并保持正确的读码框。干扰质粒转染ASTs 48h后与对照组相比，Cx43蛋白表达明显下降(P<)，细胞存活未受影响，而谷氨酸释放明显减少(P<)。结论 成功构建了抑制大鼠Cx43基因表达的siRNA载体，抑制Cx43表达可降低ASTs谷氨酸的基础释放。
[关键词]连接蛋白43；siRNA；星形胶质细胞；谷氨酸
[中图法分类号]R394-33；；R34 [文献标志码] A
Effect of silencing connexin 43 gene on release of glutamate from astrocytes in rats
Wang Lingyun, Tian Yanping , Niu jianqin,Xiao Lan (Teaching and Research Section of Histology and Embryology, Third Military Medical University, Chongqing Key Laboratory of Neurobiology，Chongqing 40038, China)
[Abstract] Objective To construct the siRNA plasmid that can effectively inhibit expression of connexin 43 (Cx43) and detect the effect of inhibited Cx43 expression on release of glutamate from astrocytes (AST). Methods siRNA oligonucleotides targeting the Cx43 gene in rats were synthesized. A . vector was constructed. After plasmids were transfected into AST by electroblotting, effect of silencing plasmids transfected into AST on expression of Cx43 protein in AST was detected by We
王凌云，田衍平,牛建钦，肖 岚（400038，重庆，第三军医大学基础部组织学与胚胎学教研室，重庆市神经科学重点实验室）
[摘要] 目的 构建能有效抑制大鼠星形胶质细胞连接蛋白43（connexin 43，Cx43）基因表达的siRNA载体，并检测抑制Cx43表达对星形胶质细胞谷氨酸释放的影响。方法 合成针对大鼠Cx43基因的siRNA寡核苷酸，。质粒通过电转方法转染原代星形胶质细胞 (astrocyte, AST）后，用免疫荧光染色及Western blot检测转染干扰质粒对大鼠ASTs Cx43蛋白表达的影响。用谷氨酸ELISA定量试剂盒检测释放至培养基中谷氨酸的浓度,并用DAPI染色进行细胞计数。结果 测序结果显示，Cx43基因的目的片段正确插入载体，并保持正确的读码框。干扰质粒转染ASTs 48h后与对照组相比，Cx43蛋白表达明显下降(P<)，细胞存活未受影响，而谷氨酸释放明显减少(P<)。结论 成功构建了抑制大鼠Cx43基因表达的siRNA载体，抑制Cx43表达可降低ASTs谷氨酸的基础释放。
[关键词]连接蛋白43；siRNA；星形胶质细胞；谷氨酸
[中图法分类号]R394-33；；R34 [文献标志码] A
Effect of silencing connexin 43 gene on release of glutamate from astrocytes in rats
Wang Lingyun, Tian Yanping , Niu jianqin,Xiao Lan (Teaching and Research Section of Histology and Embryology, Third Military Medical University, Chongqing Key Laboratory of Neurobiology，Chongqing 40038, China)
[Abstract] Objective To construct the siRNA plasmid that can effectively inhibit expression of connexin 43 (Cx43) and detect the effect of inhibited Cx43 expression on release of glutamate from astrocytes (AST). Methods siRNA oligonucleotides targeting the Cx43 gene in rats were synthesized. A . vector was constructed. After plasmids were transfected into AST by electroblotting, effect of silencing plasmids transfected into AST on expression of Cx43 protein in AST was detected by We
cx43基因干扰对大鼠星形胶质细胞谷氨酸基础释放的影响 来自淘豆网www.taodocs.com转载请标明出处.