N-cadherin通过β-catenin调控鸡胚脊髓连合纤维向对侧的投射  

N-cadherin regulates projection of spinal commissural axons via β-catenin during chicken embryonic development

作　　者：杨慈清[1] 李小英 王聪睿[1] 张必超 李晗 林俊堂 

机构地区：[1]河南省医用组织再生重点实验室,新乡453003 [2]新乡医学院生命科学技术学院,新乡453003

出　　处：《中国免疫学杂志》2015年第10期1357-1360,1374,共5页Chinese Journal of Immunology

基　　金：2013年河南省教育厅科学技术研究重点项目(13A320866);新乡医学院科研培育基金(2013QN125);新乡医学院科研培育基金(2013QN115);新乡医学院博士启动基金(505090)资助

摘　　要：目的:阐明鸡胚发育过程中神经钙黏蛋白(N-cadherin)和β-连环蛋白(β-catenin)在脊髓连合纤维发育中的作用。方法:在鸡胚发育第3天(stage22),将N-cadherin和β-catenin干扰载体通过活体电转基因技术导入脊髓,电转后继续培养3 d(stage29),取材、固定、包埋和切片。分为N-cadherin干扰组、β-catenin干扰组,β-catenin和N-cadherin共同干扰组以及转染p GU6-GFP-neo表达质粒对照组;采用荧光免疫组织化学法检测N-cadherin和β-catenin蛋白表达变化;根据GFP表达结果,观察脊髓连合纤维的投射情况。结果:在鸡胚发育过程中,在脊髓抑制N-cadherin的表达会降低β-catenin的表达;在Ncadherin抑制表达72 h后观察到脊髓连合纤维向对侧的投射产生被抑制;抑制β-catenin表达出现类似向对侧投射障碍;Ncadherin和β-catenin同时被干扰时,连合纤维投射的抑制效果相似。结论:N-cadherin参与调控脊髓连合纤维的发育过程,其部分作用机制可能是通过下调β-catenin的表达实现的。Objective：To explore the role of N-cadherin and β-catenin in the formation of spinal commissural axon projection during chicken embryonic development.Methods：Fertilized eggs were cultured for three days（stage22）,N-cadherin orβ-catenin inter-ference plasmid was injected into the neural tube and in vivo electroporation was performed.Three days after the electroporation, embryos were collected,fixed with 4%PFA,embeded with OCT,and cut into frozen sections.Four groups （ knockdown of N-cadherin orβ-catenin or both of them,and control） were included in this study.Immunohistochemistry method was used to analyze the protein expression result of N-cadherin or β-catenin.The changes of spinal commissural projections were observed with GFP fluorescence.Results：During chicken embryonic development,knockdown of N-cadherin inhibited the expression of β-catenin in the spinal cord.The commissural nerve fibers projecting to the contralateral side of the spinal cord was impaired after knockdown of N-cadherin or β-catenin;this phenotype was similar after knocking down both of them.Conclusion： N-cadherin is implicated in the formation of spinal commissural projection in the developing spinal cord,possibly viaβ-catenin.

关 键 词：鸡胚 神经钙黏蛋白 Β-连环蛋白 神经纤维 

分 类 号：Q954.48[生物学—动物学]