摘要
为探讨新基因ＨＣＹ－２与神经管畸形表型的关系及明确其是否具有基因毒性作用，应用脂质体介导基因转移技术将构建的含有全长ＨＣＹ－２ ｃＤＮＡ（２０１４ｂｐ）的真核表达载体转入新生鸡受精卵，培养３．５天后观察其对神经管闭合时期胚胎的整体分子生物学效应，利用Ｗｅｓｔｅｒｎ 印迹和免疫组化等技术研究在整体环境下ＨＣＹ－２蛋白的表达和分布情况。结果显示转ＨＣＹ－２基因（５～２０μｇ）可诱导早期鸡胚发生神经管畸变，而空载质粒组胚胎发育正常；Ｗｅｓｔｅｒｎ 印迹和免疫组化分析发现有ＨＣＹ－２蛋白大量表达，主要分布于胚脑组织中，显著高于空载质粒对照组；透射电镜观察发现转ＨＣＹ－２基因后，胚脑细胞超微结构受损和出现呈弥散分布的凋亡细胞。结论是ＨＣＹ－２基因对神经管闭合时期的胚胎神经系统发育具有明显基因毒性作用。
In order to explore the relationship between novel gene HCY 2 and neural tube defects (NTDs) and to investigate whether HCY 2 gene was genotoxic, a mammalian expression vector containing HCY 2 cDNA(2014 bp) was transferred into chick fertilized ovum by using lipofectin reagent, then the biological effects of HCY 2 gene on embryos in culture for 3.5 days were observed. Western blotting and immunohistochemistry were used to analyse the expression and distribution of HCY 2 after gene being transferred. HCY 2 gene (5～20μg/chick ovum) could induce NTDs in organogenesis period. Western blotting and immunohistochemical staining showed that HCY 2 protein could be greatly expressed by day 3.5, the time which NTDs developped, with stronger staining at the site of embryonic brain as compared with controls. Under electron microscope, it was observed that HCY 2 gene could damage embryonic cellular ultrastructures and cause excessive apoptosis of neurocytes. These findings suggested that HCY 2 gene with its product was dysmorphogenic to the neural tube, might be a genotoxic factor or teratogenic agent and should be responsible for chick NTDs and probably human NTDs as well.
出处
《卫生研究》
CAS
CSCD
北大核心
1999年第6期350-352,共3页
Journal of Hygiene Research
基金
国家"863"计划基金国家自然科学基金
