铅对斑马鱼胚胎发育过程Fgf3基因表达的影响  

Effect of lead exposure on gene expression of Fgf3 in zebrafish embryonic development

作　　者：贾聪聪[1] 林林[1] 刘妮娅[1] 张小晶[1] 张佳佳[1] 杨新军[1] 黄陈平[1] 

机构地区：[1]温州医学院环境与公共卫生学院,325035

出　　处：《中华劳动卫生职业病杂志》2012年第10期730-734,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金项目（30972509）

摘　　要：目的研究铅暴露对斑马鱼胚胎发育过程Fgf3基因表达的影响，以探讨铅的胚胎发育毒性机制。方法野生型AB品系斑马鱼胚胎暴露醋酸铅浓度分别为0、0．1、0．5、2．5和12．5μmol／L，提取各组8、12、16、24、36、48及72hpf（受精后小时）斑马鱼胚胎总RNA，实时荧光PCR检测Fgf3基因定量表达；制备Fgf3基因RNA探针，整胚原位杂交方法检测Fgf3基因空间性表达。结果在胚胎发育全程中，各组Fgf3表达高峰均在12hpf（与其他时段比较，P〈0．01），随着铅暴露浓度的升高，Fgf3 mRNA平均表达量也逐渐增加，以对照组堙移mRNA表达量为基准，各铅暴露组（0．1、0．5、2．5和12．5μmol／L）相对表达量分别为1．02±0．24、1．05±0．26、1．22±0．46和1．25±0．38，其中2．5和12．5μmol/L铅浓度组Fgf3表达量均显著高于对照组（P〈0．05）；整胚原位杂交结果显示Fgf3主要表达部位是胚胎早期的头、尾部和胚胎中后期的中脑、鳍芽及咽弓部，在胚胎发育至12hpf时Fgf3阳性杂交信号区域及强度最为明显，但各铅暴露组与对照比较，在Fgf3表达部位及强度上未见明显差异。结论胚胎期铅暴露能引起Fgf3表达水平上调，改变了胚胎发育过程Fgf3正常表达规律，其可能与铅胚胎发育毒性有关。Objective To investigate the effect of lead exposure on the gene expression of fibrohlast growth factor 3 （Fgf3） in zebrafish embryonic development and the mechanism of lead-induced embryonic developmental toxicity. Methods The embryos of zebrafish （wild types A and B） were exposed to lead acetate （PbAc） at the doses of 0, 0.1, 0.5, 2.5, and 12.5 μmol/L separately. Total RNA was extracted from each treatment group of zebrafish embryos at 8, 12, 16, 24, 36, 48, and 72 hours post fertilization （hpf）. The total mRNA expression of Fgf3 was measured by real-time quantitative PCR. The spatial expression of Fgf3 in zebrafish embryos was determined by whole-mount in situ hybridization using synthesized Fgf3 RNA probe. Results The mRNA expression of Fgf3 in each group peaked at 12 hpf （P〈0.01）. With the increase in PbAc concentration, the mRNA expression of Fgf3 rose. Compared with the mRNA expression level of Fgf3 in the control group, the relative mRNA expression levels of Fgf3 in the 0.1, 0.5, 2.5, and 12.5 μmol/L PbAc exposure groups were 1.02±0.24, 1.05±0.26, 1.22±0.46, and 1.25±0.38, respectively, and the 2.5 and 12.5 μmol/L PbAc exposure groups showed significantly higher Fgf3 expression than the control group （P〈0.05）. The whole-mount in situ hybridization results showed that Fgf3 expression occurred mainly in the head and tail in the early stage of embryonic development and in the midbrain, fin bud, and pharyngeal arch in the middle/late stage of embryonic development; there were the most significant regions and intensities of positive hybridization signals at 12 hpf; but no significant differences were found between the control group and exposure groups in the location and intensity of Fgf3 expression Conclusion Lead exposure can result in the upregulation of Fgf3 expression in zebrafish embryonic development, which might contribute to lead-induced embryonic developmental toxicity.

关 键 词：铅 成纤维细胞生长因子3 胚胎发育 基因表达 

分 类 号：R346[医药卫生—基础医学]