KLF7b对斑马鱼胚胎发育的影响

Effects of KLF7b on embryo development of zebrafish

目的:探讨KLF7b对斑马鱼胚胎发育的影响。方法:体外转录野生型KLF7b m RNA、KLF7b乙酰化位点突变体m RNA;设计合成特异性KLF7b反义吗啉代寡核苷酸(morpholino oligonueleitide,MO);在1~2细胞期注射到斑马鱼胚胎体内以实现KLF7b的过表达、位点突变与敲降;观察胚胎表型并利用定量PCR检测发育相关基因的表达变化,钙黄绿素染色观察斑马鱼骨骼发育情况。结果:过表达KLF7b对斑马鱼胚胎的发育无明显影响;敲降KLF7b或注射乙酰化位点突变的KLF7b后,斑马鱼胚胎出现心包积液、体轴和头部发育的异常,肌肉发育标志基因表达下调。结论:KLF7b可通过乙酰化修饰影响斑马鱼的发育。

Objective：To explore the effects of KLF7 b on embryo development of zebrafish. Methods：Wild-type KLF7 b m RNA and KLF7 b acetylated site mutant m RNA were transcripted in vitro. Specific m RNA and morpholino oligonueleitides（MOs）were used to overexpress or site mutate or knockdown KLF7 b by microinjection at 1~2 cells stage. The phenotypes of embryos were observed at different stages and the expressions of marker genes were tested by quantitative RT-PCR. The bone development of zebrafish was detected by calcein staining. Results：KLF7 b overexpression had no effects on embryo development of zebrafish. KLF7 b knockdown or acetylated site mutation caused embryonic abnormalities including pericardial effusion,the developmental malformations of somite and head. The marker genes of muscle were down-regulated. Conclusion：KLF7 b may affect the development of zebrafish via its own acetylation.