基础医学与临床 ›› 2009, Vol. 29 ›› Issue (6): 561-565.

• 研究论文 •    下一篇

α-突触核蛋白磷酸化参与小鼠多巴胺能神经元的保护作用

刘琦 段春礼 吴波 范春香 张韬 赵焕英 赵春礼 杨慧   

  1. 首都医科大学 北京神经科学研究所 首都医科大学神经科学研究所 首都医科大学北京神经科学研究所 首都医科大学北京神经科学研究所 首都医科大学北京神经科学研究所 北京神经再生与修复研究重点实验室
  • 收稿日期:2008-09-11 修回日期:2009-02-23 出版日期:2009-06-25 发布日期:2009-06-25
  • 通讯作者: 杨慧

α-Synuclein phosphorylation takes part in mouse dopaminergic neuron protection

Qi LIU, Chun-li DUAN, Bo WU, Chun-xiang FAN, Tao ZHANG, Huan-ying ZHAO, Chun-li ZHAO, Hui YANG   

  1. Beijing Institute for Neuroscience, Capital Medical University Beijing Institute for Neuroscience, Capital Medical University Beijing Institute for Neurosciences, Capital Medical University Beijing Institute for Neurosciences, Capital Medical University Beijing Institute for Neuroscience and Beijing Center for Neural Regeneration and Repairing, Capital Medical University
  • Received:2008-09-11 Revised:2009-02-23 Online:2009-06-25 Published:2009-06-25
  • Contact: Hui YANG

摘要: 目的 研究α-突触核蛋白(α-SYN) 129位点磷酸化修饰是否具有神经元保护作用。方法 应用反转录病毒感染小鼠多巴胺能神经元细胞MN9D,通过实时定量PCR检测各组细胞内α-SYN过表达情况,并通过免疫细胞化学方法检测野生型α-SYN (WT/α-SYN)及129位点磷酸化α-SYN (S129D/α-SYN)过表达后α-SYN在细胞内聚集情况,应用CCK-8检测细胞活力,观察WT/α-SYN及S129D/α-SYN对MN9D的细胞毒性,从而明确S129D/α-SYN对多巴胺能神经元的影响。 结果 实时定量PCR结果显示,WT/α-SYN及S129D/α-SYN在MN9D细胞内均过表达(P<0.01)。WT/α-SYN过表达组细胞活力明显下降(P<0.01),共聚焦显微镜观察未见明显的α-SYN聚集;S129D/α-SYN过表达组细胞内可观察到明显的α-SYN聚集体,同WT/α-SYN组相比细胞活力有明显提高(P<0.01)。结论 129位点丝氨酸的磷酸化修饰在一定程度上减轻了WT/α-SYN过表达所引起的细胞毒性。

关键词: α-突触核蛋白, 磷酸化α-突触核蛋白, 神经细胞保护, 多巴胺神经元

Abstract: Objective To study the role of phosphorylation at Serine 129 in regulating the neurotoxicity of α-synuclein. Methods Wild type and phosphomimic mutant α-synuclein genes were over-expressed in mouse dopaminergic cells MN9D using retrovirus. The cell viability was examined using CCK-8 assay and the cell morphology was observed with immunofluorescence microscope. Results The result of real time PCR showed that WT/α-SYN and S129D/α-SYN genes were overexpressed in MN9D as compared to uninfected MN9D and vector control group. The cell viability was obviously reduced when over-expressing both WT and phosphorylated α-synuclein genes. And the viability of cells over-expressing wild type α-synuclein was much lower than that of the cells over-expressing phosphorylated α-synuclein. Insoluble α-synuclein aggregates were observed with confocal microscope in the MN9D cells over-expressing phosphorylated α-synuclein. Conclusion Over-expressing phosphorylated α-synuclein may lead to α-synuclein aggregated in MN9D cells. Phosphorylated at 129 Ser may protect the cells from toxicity induced by over expressing α-synuclein.

Key words: a-synuclein, phosphorylated α-synuclein, neurocyte protection, dopaminergic cells