鸡胚发育过程敲减神经型钙粘蛋白表达对视顶盖神经元迁移及神经丝蛋白的影响

doi:10.3969/j.issn.0529-1356.2013.05.005

解剖学报 ›› 2013, Vol. 44 ›› Issue (5 ): 612-615.doi: 10.3969/j.issn.0529-1356.2013.05.005

鸡胚发育过程敲减神经型钙粘蛋白表达对视顶盖神经元迁移及神经丝蛋白的影响

杨慈清^1,2,3 李琼²付苏雷¹ 李晗¹ 范义峰³ 郭志坤² 林俊堂^1,2*

1. 新乡医学院生命科学技术学院细胞工程教研室， 2. 河南省医用组织再生重点实验室，河南新乡 453003; 3. 新乡市驼人医疗器械有限公司，河南新乡 453403

收稿日期:2012-12-14 修回日期:2013-04-03 出版日期:2013-10-06 发布日期:2013-10-06
通讯作者: 林俊堂 E-mail:linjuntang2002@yahoo.co.uk
基金资助:
国家自然科学基金资助项目

Effect of knocking down N-cadherin expression on the tectum neuronal migration and neurofilament expression during chick embryo development

YANG Ci-qing^1,2,3 LI Qiong²FU Su-lei¹ LI Han¹ FAN Yi-feng³GUO Zhi-kun²LIN Jun-tang ^1,2*

1. Department of Cell Engineering, College of Life Science and Technology, Xinxiang Medical University ; 2.Key Laboratory of Henan Province for Medical Tissue Regeneration, Xinxiang Medical University， He’nan Xinxiang 453003，China;3. Xinxiang Tuoren Medical Devices Co. Ltd., He’nan Xinxiang 453003，China


Received:2012-12-14 Revised:2013-04-03 Online:2013-10-06 Published:2013-10-06

摘要/Abstract

摘要：

目的阐明鸡胚发育过程神经型钙黏蛋白（N-cadherin）在中枢神经系统中的作用及其对神经丝蛋白(NF)表达的影响。方法鸡胚正常培养到第3天（E3），采用鸡胚带壳开窗培养方法，取出3～4ml蛋清，开窗培养至E5，将N-cadherin干扰载体pGPU6-GFP-neo-N-cadherin-shRNA通过活体电转基因技术导入视顶盖，电转后继续培养到E12，每组收集3个胚胎，取材，固定，包埋，切片后进行荧光免疫组织化学分析相关蛋白的表达。结果鸡胚发育过程，敲减N-cadherin表达影响神经元迁移，被敲减的细胞主要停留在室管膜区，在N-cadherin受到抑制表达的区域，NF的表达也受到抑制，其表达明显减弱。结论 N-cadherin的抑制表达可导致神经元的迁移发生紊乱，影响NF的表达。

关键词: 胚胎发育, 视顶盖, 神经型钙黏蛋白, 神经丝蛋白, 活体电转, 鸡胚

Abstract:

Objective To clarify the role and function of the N-cadherin and its impact on the expression of neurofilaments in central nervous system during the chick embryo development. Methods Fresh eggs were cultured in normal condition for three days (E3), and 3-4ml of egg white was removed from egg, then continuing culture to E5. N-cadherin interference plasmids (pGPU6-GFP-neo-N-cadherin-shRNA) was injected into the optic tectum and then handled by electroporationin vivotechnology. Tissues were obtained at E12 and from each group three embryos were collected, fixed with 4%PFA, embeded with OCT, and cut into frozen sections. Fluorescence immunohistochemistry method was used to analyses the different protein expression. Results Expression of N-cadherin was inhibited, which caused neuronal migration disorder. The expression of NF was also suppressed. Conclusion Inhibition of N-cadherin expression can lead to neuronal migration disorder, affecting the expression of NF.

Key words: Embryonic development, Optic tectum, Neural cadherin, Neurofilament, Electroporationin vivo, Chick embryo

杨慈清李琼付苏雷李晗范义峰郭志坤林俊堂* . 鸡胚发育过程敲减神经型钙粘蛋白表达对视顶盖神经元迁移及神经丝蛋白的影响[J]. 解剖学报, 2013, 44(5 ): 612-615.

YANG Ci-qing LI Qiong FU Su-lei LI Han FAN Yi-feng GUO Zhi-kun LIN Jun-tang*. Effect of knocking down N-cadherin expression on the tectum neuronal migration and neurofilament expression during chick embryo development[J]. AAS, 2013, 44(5 ): 612-615.

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鸡胚发育过程敲减神经型钙粘蛋白表达对视顶盖神经元迁移及神经丝蛋白的影响

Effect of knocking down N-cadherin expression on the tectum neuronal migration and neurofilament expression during chick embryo development

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