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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
杨慈清 李琼 付苏雷 李晗 范义峰 郭志坤 林俊堂*
解剖学报 ›› 2013, Vol. 44 ›› Issue (5) : 612-615.
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
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。
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 electroporationin vivotechnology. 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.
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
Embryonic development / Optic tectum / Neural cadherin / Neurofilament / Electroporationin vivo / Chick embryo
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