CAJ | 학술논문

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pGU6－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N－cadherin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；N－cadherin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。
목적：천명계배발육과정중신경개점단백（ N－cadherin）화β－련배단백（β－catenin）재척수련합섬유발육중적작용。방법：재계배발육제3천（stage22），장N－cadherin화β－catenin간우재체통과활체전전기인기술도입척수，전전후계속배양3 d（stage29），취재、고정、포매화절편。분위N－cadherin간우조、β－catenin간우조，β－catenin화N－cadherin공동간우조이급전염pGU6－GFP－neo표체질립대조조；채용형광면역조직화학법검측N－cadherin화β－catenin단백표체변화；근거GFP표체결과，관찰척수련합섬유적투사정황。결과：재계배발육과정중，재척수억제N－cadherin적표체회강저β－catenin적표체；재N－cadherin억제표체72 h후관찰도척수련합섬유향대측적투사산생피억제；억제β－catenin표체출현유사향대측투사장애；N－cadherin화β－catenin동시피간우시，련합섬유투사적억제효과상사。결론：N－cadherin삼여조공척수련합섬유적발육과정，기부분작용궤제가능시통과하조β－catenin적표체실현적。
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.