Li Y, Li Z, Chen X, Meng Z
National Center for Maternal and Infant Health Care of China, Beijing Medical University, Beijing 100083, China.
Wei Sheng Yan Jiu. 1999 Nov;28(6):350-2.
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 (5-20 micrograms/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 developed, 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.
为了探究新基因HCY - 2与神经管缺陷(NTDs)之间的关系,并研究HCY - 2基因是否具有遗传毒性,利用脂质体转染试剂将含有HCY - 2 cDNA(2014 bp)的哺乳动物表达载体导入鸡受精卵,然后观察HCY - 2基因对培养3.5天的胚胎的生物学效应。基因转染后,采用蛋白质免疫印迹法和免疫组织化学法分析HCY - 2的表达和分布。HCY - 2基因(5 - 20微克/鸡胚)可在器官发生期诱导神经管缺陷。蛋白质免疫印迹法和免疫组织化学染色显示,在神经管缺陷发生的3.5天时,HCY - 2蛋白大量表达,与对照组相比,胚胎脑部位染色更强。在电子显微镜下观察到,HCY - 2基因可破坏胚胎细胞超微结构并导致神经细胞过度凋亡。这些发现表明,HCY - 2基因及其产物对神经管具有致畸作用,可能是一种遗传毒性因子或致畸剂,应对鸡的神经管缺陷以及可能的人类神经管缺陷负责。