α-突触核蛋白寡聚体抑制大鼠原代培养神经元突起早期生长

doi:10.3969/j.issn.1006-7795.2014.05.013

首都医科大学学报 ›› 2014, Vol. 35 ›› Issue (5): 587-591.doi: 10.3969/j.issn.1006-7795.2014.05.013

• α-突触核蛋白改变对神经元的影响 • 上一篇下一篇

α-突触核蛋白寡聚体抑制大鼠原代培养神经元突起早期生长

王鹏^1,2, 李昕¹, 陈予东¹, 杨巍巍¹, 李旭冉¹, 于顺¹

1. 首都医科大学宣武医院神经生物学研究室, 北京 100053;
2. 北华大学基础医学院人体解剖学教研室, 吉林吉林 132013

收稿日期:2014-07-21 出版日期:2014-10-21 发布日期:2014-10-20
通讯作者: 于顺 E-mail:yushun103@yahoo.com.cn
基金资助:
国家重点基础研究发展计划（2011CB504101），国家自然科学基金（81071014，81371200），国家科技支撑计划课题（2012BAI10B03），首都卫生发展科研专项课题（2011-4001-01），北京市自然科学基金（7122035）。

Effect of α-synuclein oligomer inhibit neurite outgrowth of rat primary cultured neuron

Wang Peng^1,2, Li Xin¹, Chen Yudong¹, Yang Weiwei¹, Li Xuran¹, Yu Shun¹

1. Department of Neurobiology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;
2. Department of Human Anatomy, School of Basic Medical Sciences, Beihua University, Jilin 132013, Jilin Province, China

Received:2014-07-21 Online:2014-10-21 Published:2014-10-20
Supported by:
This study was supported by National Basic Research Program (973 Program) of China (2011CB504101), National Natural Science Foundation of China (81071014, 81371200), National Science and Technology Support Program (2012BAI10B03), Capital Health Research and Development Special Fund (2011-4001-01), Natural Science Foundation of Beijing (7122035).

摘要/Abstract

摘要：

目的观察人α-突触核蛋白（α-synuclein，α-Syn）和其形成的寡聚体对大鼠原代培养神经元突起生长的影响，明确α-Syn的生理功能，探讨其寡聚化对神经元的毒性及其在帕金森病发病机制中的作用。方法制备α-Syn的寡聚体，向分组培养的大鼠脑皮质神经元培养基中添加，培养1、2和4 h后固定，观察对神经元突起生长的影响。神经元的突起以成像显微镜观察测量，Western blotting法、免疫荧光法实验进行鉴定。结果添加α-Syn寡聚体组的神经元培养至1、2和4 h时，其突起的平均长度均小于对照组和添加α-Syn单体组（P<0.05）。增加α-Syn寡聚体浓度，随浓度增高神经元突起的平均长度与对照组差异越大（P<0.01）。Western blotting法和免疫荧光实验明确了外源性α-Syn寡聚体可以从培养基进入到神经元内。结论 α-Syn寡聚体在原代神经元生长初期对其突起生长具有抑制作用，这一现象可能与其在帕金森病发病机制中的作用有关。

关键词: α-突触核蛋白, 寡聚体, 原代神经元培养, 突起生长, 帕金森病

Abstract:

Objective To investigate the effect of oligomerization of α-synuclein on the neurite outgrowth of brain neurons and indicate its function, explore the mechanism of Parkinson's disease. Methods Neurons isolated from the neocortex of newborn SD rats were cultured and corresponding proteins were added into the culture medium to observe the effect of the protein on the neurite outgrowth of the neurons. Added α-Syn oligomer, α-synuclein monomer into the culture system of neural cells, and compared the different promoting effect of neurite outgrowth with that of control groups. Western blotting assays and immunofluorescence staining assay were used to confirm whether the result is special. Observed with inverted phase contrast microscope,and photographs were applied to analyze with software.Results The mean neurite length of the neurons treated by α-Syn oligomer was significantly shorter than that of the control group neurons (P<0.05). But the length of neurons treated by α-Syn monomer is longer than that of the control group (P<0.05). Increased concentration of α-synuclein oligomer can enhance this effect. The result of Western blotting and Immunofluorescence staining assay showed that α-synuclein oligomer can enter the neural cell and inhibit the neurite outgrowth. Conclusion α-Synuclein can inhibit neurite outgrowth of primary cultured neuron and this is a dose-dependent effect.

Key words: α-synuclein, oligomer, primary neuronal culture, neurite outgrowth, Parkinson’s disease

中图分类号:

王鹏, 李昕, 陈予东, 杨巍巍, 李旭冉, 于顺. α-突触核蛋白寡聚体抑制大鼠原代培养神经元突起早期生长[J]. 首都医科大学学报, 2014, 35(5): 587-591.

Wang Peng, Li Xin, Chen Yudong, Yang Weiwei, Li Xuran, Yu Shun. Effect of α-synuclein oligomer inhibit neurite outgrowth of rat primary cultured neuron[J]. Journal of Capital Medical University, 2014, 35(5): 587-591.

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α-突触核蛋白寡聚体抑制大鼠原代培养神经元突起早期生长

Effect of α-synuclein oligomer inhibit neurite outgrowth of rat primary cultured neuron

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