鱼藤素可影响斑马鱼胚胎Wnt信号通路相关基因的表达

背景：前期研究发现,在斑马鱼早期发育的不同时期,经过一定浓度鱼藤素处理的胚胎表现出不同程度的发育延缓和表型异常。目的：观察斑马鱼早期胚胎发育过程中鱼藤素对wnt信号通路相关基因表达的影响。方法：收集孵育至shield期的斑马鱼胚胎于6孔板中,加入0.6 μmol/L浓度的鱼藤素溶液,避光处理,28 ℃培养箱中孵养。用0.1%二甲基亚砜溶液作为对照组,相同条件培养。在受精后6,8,10,12,14,24 h时用正置荧光显微镜拍照观察鱼藤素处理后斑马鱼胚胎发育表型。实时荧光定量核酸扩增检测鱼藤素作用前后目的基因表达水平的变化。结果与结论：shield期处理的斑马鱼胚胎在bud期停止生长;wnt信号通路上的wnt9a,wnt10b和axin2以及与wnt信号具有密切联系的snail1b,Jnk2表达发生显著变化,其中wnt9a,wnt10b,axin2,snail1b表达下调,Jnk2表达上调。结果提示鱼藤素对斑马鱼体内wnt信号具有负调节作用。

Deguelin regulates the expression of Wnt signaling pathway related genes in Zebrafish embryo

BACKGROUND: Previous studies found that in different periods of early development of zebrafish, embryos treated with certain concentration of deguelin show varying degrees of developmental delay and abnormal phenotype. OBJECTIVE:To observe the effect of deguelin on expression of Wnt signaling pathway related genes in Zebrafish embryo during early embryo development. METHODS: Zebrafish embryos that incubated to the shield period were collected and put into the 6-well plate, 0.6 μmol/L deguelin solution were added and treated in dark, and then cultured in the incubator under 28 ℃. Zebrafish embryos treated with 0.1% dimethylsulfoxide solution were as control group, and cultured in the same conditions. At 6, 8, 10, 12, 14 and 24 hours after fertilization, phenotype of Zebrafish embryos was observed with upright fluorescence microscope. The expression levels of target genes before and after treatment were tested by quantitative real time PCR.RESULTS AND CONCLUSION:The embryos treated by deguelin on shield stage stop growing and stay at bud stage; the expressions of wnt9a, wnt10b, axin2 as well as snail1b and Jnk2 related with Wnt signal were changed significantly, among them, the expression of wnt9a, wnt10b, axin2 and snail 1b were decreased, while the expression of Jnk2 was increased. Deguelin has a negative effect on Zebrafish in vivo Wnt signal.