突变型α-突触核蛋白对大鼠原代培养神经元突起生长作用研究

doi:10.3969/j.issn.1006-7795.2012.05.015

首都医科大学学报 ›› 2012, Vol. 33 ›› Issue (5): 629-633.doi: 10.3969/j.issn.1006-7795.2012.05.015

突变型α-突触核蛋白对大鼠原代培养神经元突起生长作用研究

王鹏¹, 许洁², 李昕³, 何欣¹, 李尧华³, 于顺³

1. 北华大学基础医学院人体解剖学教研室,吉林 132013;2. 吉林化工学院理学院,吉林 132022;3. 首都医科大学宣武医院老年病研究所神经生物学研究室,北京 100053

收稿日期:2012-04-25 修回日期:1900-01-01 出版日期:2012-10-21 发布日期:2012-10-21
通讯作者: 于顺

Enhancing effect of mutant α-synuclein on neurite outgrowth of rat primary cultured neuron

WANG Peng¹, XU Jie², LI Xin³, HE Xin¹, LI Yao-hua³, YU Shun³

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. Department of Neurobiology, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Diseases of Ministry of Education, Beijing 100053, China

Received:2012-04-25 Revised:1900-01-01 Online:2012-10-21 Published:2012-10-21

摘要/Abstract

摘要： 目的观察人α-突触核蛋白(α-synuclein,α-Syn)和其突变体A30P和A53T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,揭示突变体A30P和A53T在帕金森病的发病机制中的作用。方法取大鼠大脑皮质神经元分组培养,在细胞外添加A30P、A53T和α-Syn,培养1 h、2 h和4 h后固定,比较A30P、A53T与α-Syn对神经元突起生长的影响。神经元突起以成像显微镜观察测量,Western blotting法、免疫荧光法、单克隆抗体阻断实验鉴定各蛋白作用的特异性。结果添加α-Syn组的神经元培养至1、2和4 h时,其突起的平均长度大于对照组和添加A30P、A53T组 (P<0.05)。A30P、A53T组和对照组的神经元突起长度差异无统计学意义(P>0.05)。增加蛋白的含量,浓度越高,α-Syn组神经元突起的平均长度与添加A30P和A53T组差异越大(P<0.01)。Western blotting和免疫荧光实验明确了外源性α-Syn可以从培养基进入到神经元内,并均匀分布在胞体和突起。而A30P和A53T组,并未发现。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,突变体A30P和A53T,无促神经元突起生长作用,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: 突触核蛋白, 原代神经元培养, 突起生长, 帕金森病

Abstract: Objective To investigate the effect of A53T and A30P 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 Wistar 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. A53T, A30P and α-synuclein were added into the culture system of neural cells, and the different promoting effect of neurite outgrowth was compared with that of control groups. Western blotting assays and immunofluorescence staining assay were used to confirm whether the result is special. Inverted phase contrast microscope,and photographs were applied to analyze the results with software. Results The mean neurite length of the neurons treated with α-Syn was significantly longer than that of A53T and A30P group neurons(P<0.05). There was no significant difference between A53T and A30P groups with control group(P>0.05). Increased concentration of α-synuclein could enhance this effect. The result of Western blotting and immunofluorescence staining assay showed that α-synuclein could enter the neural cell and enhance the neurite outgrowth. Blockade with monoclonal antibody 3D5 showed that the blocking of α-Syn could significantly inhibit the effect (P>0.05). Conclusion α-Synuclein can enhance neurite outgrowth of primary cultured neuron in a dose-dependent manner. But its mutants A53T and A30P did not show such effect.

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

中图分类号:

R 74

王鹏;许洁;李昕;何欣;李尧华;于顺. 突变型α-突触核蛋白对大鼠原代培养神经元突起生长作用研究[J]. 首都医科大学学报, 2012, 33(5): 629-633.

WANG Peng;XU Jie;LI Xin;HE Xin;LI Yao-hua;YU Shun. Enhancing effect of mutant α-synuclein on neurite outgrowth of rat primary cultured neuron[J]. Journal of Capital Medical University, 2012, 33(5): 629-633.

[1]	粟文婷, 於佳楠, 贾军, 王晓民. 电针对帕金森病模型小鼠纹状体背侧的c-Fos表达的选择性调节[J]. 首都医科大学学报, 2021, 42(6): 986-992.
[2]	高歌, 查旭, 刘伟进, 杨慧. 129位丝氨酸磷酸化α-突触核蛋白DELFIA检测方法的建立及初步应用[J]. 首都医科大学学报, 2021, 42(5): 776-782.
[3]	王梦月, 赵鑫, 曹枭, 王可, 贾军. 帕金森病小鼠模型自主活动状态下的运动启动分析[J]. 首都医科大学学报, 2021, 42(4): 553-558.
[4]	袁惠莉, 宫晓丽, 范征, 梁志刚, 王晓民. 远志皂苷对脂多糖诱导的帕金森病模型大鼠的作用及机制研究[J]. 首都医科大学学报, 2020, 41(6): 914-922.
[5]	宋启晗, 李旭冉, 李昕, 杨巍巍, 李伟, 于顺. 不同亚型的帕金森病及多系统萎缩患者血浆α-突触核蛋白寡聚体浓度分析[J]. 首都医科大学学报, 2020, 41(2): 212-216.
[6]	丁晖, 蔡彦宁, 陈彪. 灵芝提取物对MPP⁺诱导的Neuro-2a细胞自噬的影响[J]. 首都医科大学学报, 2019, 40(4): 596-601.
[7]	张辉, 马惠清, 王森茂, 王晓娟. 银杏叶提取物对帕金森病模型大鼠脑内神经炎性反应的影响[J]. 首都医科大学学报, 2019, 40(3): 439-442.
[8]	李杰, 谢秀娟, 宋炜, 潘维花, 杨巍巍, 于顺. 帕金森病患者血浆增强α-突触核蛋白细胞毒性及其机制研究[J]. 首都医科大学学报, 2019, 40(2): 244-250.
[9]	宫晓丽, 刘梦茹, 王乐, 刘玚, 张婷, 孙英, 王晓民. α-突触核蛋白在小胶质细胞炎性反应和吞噬功能中的作用研究[J]. 首都医科大学学报, 2018, 39(5): 693-698.
[10]	刘梦茹, 宫晓丽, 王乐, 刘玚, 张婷, 王晓民. 脂多糖对肠道α-突触核蛋白表达的影响[J]. 首都医科大学学报, 2018, 39(4): 557-562.
[11]	罗斌斌, 胡亚松, 岳峰, 陈彪. MPTP诱导的食蟹猴偏侧帕金森病模型运动行为学的长期动态评价研究[J]. 首都医科大学学报, 2018, 39(2): 217-222.
[12]	卢勇泉, 陈敏, 高歌, 段春礼, 鲁玲玲, 杨慧. TRPC3参与α-突触核蛋白引起的线粒体损伤[J]. 首都医科大学学报, 2017, 38(6): 857-862.
[13]	陈敏, 于文娇, 李昕, 杨巍巍, 李旭冉, 于顺. α-突触核蛋白通过上调内吞蛋白Rab5B促进海马神经元膜表面NMDA受体内在化[J]. 首都医科大学学报, 2017, 38(6): 863-867.
[14]	于文娇, 杨巍巍, 李昕, 李旭冉, 陈敏, 于顺. α-突触核蛋白通过Rab5B下调海马神经元膜表面NMDA受体含量及其介导的Ca²⁺内流和内向电流[J]. 首都医科大学学报, 2017, 38(6): 868-873.
[15]	杨巍巍, 李昕, 李旭冉, 于顺. 食蟹猴脑与结肠中寡聚化和磷酸化α-突触核蛋白的增龄性变化的相关性分析[J]. 首都医科大学学报, 2017, 38(6): 874-878.

突变型α-突触核蛋白对大鼠原代培养神经元突起生长作用研究

Enhancing effect of mutant α-synuclein on neurite outgrowth of rat primary cultured neuron

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价