J4 ›› 2012, Vol. 38 ›› Issue (6): 1101-1105.

• 基础研究 • 上一篇    下一篇

α-突触核蛋白功能片段对原代培养大鼠神经元突起的促生长作用

王 鹏1|许 洁2|李 昕3|于 顺3|何 欣1   

  1. 1. 北华大学基础医学院人体解剖学教研室|吉林 吉林 132013;2.吉林化工学院理学院|吉林 吉林 132022;
    3.首都医科大学宣武医院 北京老年病研究所神经生物学研究室|北京 100053
  • 收稿日期:2012-06-07 发布日期:2012-11-28
  • 通讯作者: 何 欣(Tel:0432-64608003; E-mail:hexin8040@163.com) E-mail:hexin8040@163.com
  • 作者简介:王 鹏(1977-)|男|吉林省吉林市人|讲师|医学硕士|主要从事人体解剖学基础理论和老年病神经生物学研究。
  • 基金资助:

    吉林省教育厅“十二五”科学技术研究项目资助课题(吉教科合字[2012]第137号)

Growth-promoting effect of α-Synuclein function fragment on neurites of primary cultured neurons of rats


WANG Peng1,XU Jie2|LI Xin3,YU Shun3,HE Xin1   

  1. 1.Department of Human Anatomy,School of Basic Medical Sciences,Beihua University,Jilin 132013,China|2.College of Science,Jilin Institute of Chemical Technology,Jilin 132022|China;3.Research Room of Neurobiology,Beijing Institute of Geriatrics,Xuanwu Hospital
    ,Capital University of Medical Sciences,Beijing 100053,China
  • Received:2012-06-07 Published:2012-11-28

摘要:

目的:研究人α-突触核蛋白(α-Syn)对原代培养大鼠神经元突起的促生长作用,阐明该蛋白质的功能片段,并探讨其作用机制。方法:获取新生Wistar大鼠大脑皮层神经元分组培养,分别加入α-Syn(添加α-Syn组)、α-Syn功能片段N端(添加N端组)、C端(添加C端组)和NAC段(添加NAC段组),对照组不添加功能片段。倒置相差显微镜观察各组神经元突起的长度,Western blotting法、免疫荧光法特异性鉴定各组α-Syn蛋白的表达水平。结果:在生长至1和2 h, 添加C端组和α-Syn组的神经元突起平均长度长于对照组(P<0.05),而添加NAC段组和N端组与对照组比较差异无统计学意义(P>0.05);生长至4 h,添加α-Syn组和C端组的神经元突起平均长度明显长于对照组(P<0.01),而添加NAC段组和N端组与对照组比较差异无统计学意义(P>0.05)。Western blotting和免疫荧光法检测,N端和C端可以进入神经元,NAC段不能进入神经元。结论:α-Syn在原代神经元生长初期对其突起生长具有促进作用,具有这一功能的片段位于C端,其机制可能是其通过胞膜进入到神经元内,促进微管蛋白聚合形成微管,加速细胞骨架的形成和轴浆转运。

关键词: 突触核蛋白;神经元;突起生长;细胞培养;帕金森病

Abstract:

Objective To study the  growth-promoting effect of α-Synuclein (α-Syn) function fragment on neurites of primary cultured neurons of rats and to clarify its physiological function fragment  and to explore its mechanism.Methods The neurons in neocortex of newborn SD rats were cultured in various groups.The α-syn (adding α-Syn group) and its function segments such as  N segment(adding N segment group),C segment (adding C  segment group) and NAC segment (adding NAC segment group)  were added into the neural cells,and no protein was added  into  control group.The lengths of neurites of neurons were observed with inverted phase contrast microscope.Western blotting  and immunofluorescence staining assay were used to confirm the expression levels of proteins in various groups.Results 1 and 2 h after culture,the average lengths of neurites in  adding C segment group and adding α-Syn group were longer than that in control group (P<0.05);the average lengths of neuites in  adding NAC and N segment groups had no significant differences compared with control group(P>0.05). 4 h after culture,the average length of neurites in  adding C segment group was  significantly higher than that in control group (P<0.01);the average lengths of neuites in adding NAC and N segment groups had no significant difference compared with control group(P>0.05).The results of Western blotting and immunofluorescence staining assay showed  that N segment and C segment could enter into the neuron,but NAC could not enter into the neuron.Conclusion α-Syn can enhance the outgrowth of neurite  of primarily cultured neuron.The C segment of α-Syn has the potential to enhance neurite outgrowth,and its possible mechanism may be that C segment of α-Syn  can enter into the neuron,and  polymerize tubulin into microtubule,and accelerate the formation of cytoskeleton and axoplasmic transporting.

Key words: α-Synuclein, neuron;neurite outgrowth, cell culture;Parkinson’s disease

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