RT-PCR法检测神经源性分化因子mRNA在大鼠脑发育过程中的表达

doi:10.3969/j.issn.0529-1356.2014.01.005

解剖学报 ›› 2014, Vol. 45 ›› Issue (1): 26-30.doi: 10.3969/j.issn.0529-1356.2014.01.005

RT-PCR法检测神经源性分化因子mRNA在大鼠脑发育过程中的表达

郭玮刘晓艳柯荔宁李玉宇林凌徐剑文*

福建医科大学人体解剖学与组织学胚胎学系，神经生物中心，福州 350108

收稿日期:2013-01-08 修回日期:2013-06-05 出版日期:2014-02-06 发布日期:2014-02-06
通讯作者: 徐剑文 E-mail:fjxjw@126.com
基金资助:
福建省教育厅课题(JA07109);福建省教育厅课题(JA03086);福建医科大学教授学术发展基金(JS11003);其他(不属于以上基金类别的请自行输入下框)

A semi-quantitative RT-PCR analysis for NeuroD mRNA expression in brain during rat development

GUO Wei LIU Xiao-yan KE Li-ning LI Yu-yu LIN Ling XU Jian-wen*

Department of Anatomy, Histology & Embrgology, Fujian Medical University, Center for Neuroscience Research, Fuzhou 350108, China

Received:2013-01-08 Revised:2013-06-05 Online:2014-02-06 Published:2014-02-06
Contact: XU Jian-wen E-mail:fjxjw@126.com

摘要/Abstract

摘要：

目的观察大鼠不同发育时期脑内神经源性分化因子（NeuroD）mRNA表达量的变化，探讨其对神经细胞的分化和成熟发挥的作用。方法提取不同发育阶段大鼠不同脑组织中的mRNA，RT-PCR法反转录扩增NeuroD和β-actin，利用凝胶成像系统检测其相对表达量。结果 NeuroD在受孕7.5d（E7.5）时出现表达，在E10.5和E21.5时分别达到两次高峰，平均吸光度值为20437.88±598.28和14482.23±1134.49，表达水平显著高于其他各组（P<0.01），NeuroD/β-actin比值在E12.5、E18.5和E21.5分别为1.59±0.09、1.61±0.07和1.70±0.11，显著高于其他时间段（P<0.01）。结论在大鼠脑发育过程中，NeuroD mRNA的表达呈明显时间特异性,在E10.5左右大鼠各原始脑泡的形成过程中，NeuroD-mRNA明显增高；在胚胎发育的晚期，脑组织形态进一步完善和成熟的过程中NeuroD达到第二次表达高峰，与鼠脑的发育过程相符。推测NeuroD对鼠脑的早期神经细胞分化以及晚期细胞的成熟均起一定作用。

关键词: 神经源性分化因子, 碱性螺旋-环-螺旋, 神经发育, 神经分化, 反转录-聚合酶链反应, 大鼠

Abstract:

Objective To observe the NeuroD mRNA expression during rat neural system development and to investigate its possible roles in neural development and differentiation. Methods Total mRNA was abstracted from different parts of brains in different development stages with Trizol kit in rats. The mRNA of NeuroD and β-actin was reversely transcripted and amplificated by semi-quantitative reverse transcription-polymerase chain reaction method (RT-PCR). The quantity of NeuroD and β-actin mRNA expression was detected by gel imaging system. Results NeuroD mRNA existed in E7.5. The mRNA expression of NeuroD and β-actin peaked in E10.5 and E21.5, which average gray values were 20437.88±598.28 and 14482.23±1134.49, respectively. The mRNA expression of NeuroD and β-actin in E10.5 and E21.5 was significantly higher than other time points (P<0.01). The ratios of NeuroD to β-actin in E12.5,E18.5 and E21.5 were 1.59±0.09、1.61±0.07 and 1.70±0.11 respectively, which were higher than other time points(P<0.01). Conclusion The expression of NeuroD mRNA showed an apparent time-specific pattern during the development of the rat brain. The expression level of neuroD increased obviously in two phases. One was in E10.5, when the original rat brain came into being; the other was the anaphase in neural development, when brain form made the perfection, brain tissue stepped into its maturation. Thus, our results suggested the functions of NeuroD: 1. NeuroD influenced the generation and differentiation of nerve cells; 2. NeuroD played its role in the maturation of nervous system.

Key words: Neurogenic differentiation, Basic helix-loop-helix, Neuronal development, Neuronal differentiation, RT-PCR, Rat

中图分类号:

R329.21 R394.2

郭玮刘晓艳柯荔宁李玉宇林凌徐剑文*. RT-PCR法检测神经源性分化因子mRNA在大鼠脑发育过程中的表达[J]. 解剖学报, 2014, 45(1): 26-30.

GUO Wei LIU Xiao-yan KE Li-ning LI Yu-yu LIN Ling XU Jian-wen*. A semi-quantitative RT-PCR analysis for NeuroD mRNA expression in brain during rat development[J]. AAS, 2014, 45(1): 26-30.

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RT-PCR法检测神经源性分化因子mRNA在大鼠脑发育过程中的表达

A semi-quantitative RT-PCR analysis for NeuroD mRNA expression in brain during rat development

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