您的位置:山东大学 -> 科技期刊社 -> 《山东大学学报(医学版)》

山东大学学报(医学版) ›› 2209, Vol. 47 ›› Issue (6): 28-32.

• 论文 • 上一篇    下一篇

体外原代培养胎鼠大脑皮层神经元NMDAR1亚基表达的发育性变化

张道来 孙涛 谢珊珊 王玉卓 赵玲 冯玉新 辛华   

  1. 山东大学医学院细胞生物学研究所, 济南 250012
  • 收稿日期:2009-01-04 出版日期:2009-06-16 发布日期:2209-06-25
  • 通讯作者: 辛华(1950- ),女,教授,主要从事神经损伤与保护机制研究。 E-mail:xinhua@sdu.edu.cn
  • 作者简介:张道来(1984- ),男,硕士研究生,主要从事细胞生物学研究。E-mail: daolaizhang@yahoo.com.cn

Developmental expression of NMDAR1 subunit on cultured cerebral cortical neurons of fetal rats in vitro

 ZHANG Dao-Lai, SUN Tao, XIE Shan-Shan, WANG Yu-Zhuo, ZHAO Ling, FENG Yu-Xin, XIN Hua   

  1. Institute of Cell Biology, School of Medicine, Shandong University,Jinan 250012, China
  • Received:2009-01-04 Online:2009-06-16 Published:2209-06-25

摘要:

目的 探讨体外原代培养胎鼠大脑皮层神经元NMDAR1(NR1)亚基表达的发育性变化。 方法 利用体外原代培养Wistar孕14~15d胎鼠大脑皮层神经元,采用免疫荧光法鉴定神经元的纯度和NR1亚基在神经元上的定位,采用流式细胞术和免疫印迹法(Western  blot)检测不同培养时间神经元NR1亚基的表达情况 结果 体外原代培养的胎鼠大脑皮层神经元纯度均达到90%以上,免疫荧光标记显示NR1亚基主要分布在神经元胞膜及树突干膜上,流式细胞仪检测培养1、2、3、4、6、9、12d神经元NR1亚基平均荧光强度分别为4.57±0.99、8.18±1.22、13.67±1.99、20.33±1.03、26.30±2.88、31.71±2.47、28.63±1.40,Western blot显示细胞膜蛋白中NR1亚基在1~2d表达微弱,3~6d表达逐渐增高,6d以后保持稳定。  结论 体外原代培养的胎鼠大脑皮层神经元NR1亚基有发育性变化,这种变化与神经元的成熟度有关。

关键词: 大脑皮质;神经元;细胞培养;受体,N-甲基-D-天冬氦酸;大鼠,Wistar

Abstract:

Objective To investigate developmental expression of NMDAR1(NR1) subunit on cultured cerebral cortical neurons of fetal rats in vitro.  Methods  Cerebral cortical neurons were primarily cultured from pregnant 14~15d fetal rats.Purity of neurons and location of NR1 subunit were identified by immunofluorescence, and expression of NR1 subunit was determined by flow cytometry and Western blot.  Results The purity of neurons was more than 90%. Immunofluorescence showed that NR1 subunit is mainly located on the membrane of neuronic bodies and dendritic stems. Flow cytometry showed that average fluorescence intensity of NR1 subunit on 1d, 2d, 3d, 4d, 6d, 9d, and 12d was 4.57±0.99, 8.18±1.22, 13.67±1.99, 20.33±1.03, 26.30±2.88, 31.71±2.47, and 28.63±1.40,respectively. Western blot showed that the membrane protein NR1 subunit expression was weak on 1-2d, and gradually strengthened on 3-6d, then remained stably.  Conclusion Expression of NR1 subunit on cultured cerebral cortical neurons of fetal rats has developmental changes, which is related to neuronic maturity.

Key words: Cerebral cortex; Neurons; Cell culture; Receptors,N-methyl-D-aspartate; Rats, Wistar

中图分类号: 

  • Q291
[1] 张玉颖,张晾,潘杰. Tet-on系统诱导COX1基因在小鼠前脂肪细胞3T3-L1中的表达[J]. 山东大学学报(医学版), 2013, 51(5): 24-28.
[2] 沈涛1,代炜2,李妍3,许晓军1,巴根1,付勤1. 骨肉瘤细胞中SORBS1的表达和定位[J]. 山东大学学报(医学版), 2012, 50(2): 21-.
[3] 庄泳1, 李栋2, 时庆2, 汪大琨2, 蔡直锋1, 鞠秀丽1. 血小板衍生生长因子对人脐带间充质干细胞胞外基质的影响[J]. 山东大学学报(医学版), 2010, 48(9): 59-63.
[4] 张道来 孙涛 谢珊珊 王玉卓 赵玲 冯玉新 辛华. 体外原代培养胎鼠大脑皮层神经元NMDAR1亚基表达的发育性变化[J]. 山东大学学报(医学版), 2009, 47(6): 28-32.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!