Irx5a基因过表达对斑马鱼胚胎早期造血的影响

摘要

Objective To investigate the expression and influence of early hematopoietic related genes in zebrafish embryos.Method The single-cell-stage zebrafish embryos were micro-injected with iroquois homebox 5a-enhanced green fluorescent protein (Irx5a-EGFP) mRNA as experimental group,EGFP mRNA as experimental control group,wild type zebrafish as blank control group.Comparised the hematopoietic transcript factor genes with three groups by quantificational reverse transcriptionPCR (qPCR).Such as primitive hematopoiesis scl and lmo2,definitive hematopoiesis c-myb and runx1,erythroid hematopoiesis gata-1,myeloid hematopoiesis pu.1.The zebrafish embryos' time point are 9 h、12 h、18 h、24 h、30 h post fertilization (pf).Results From the real-time fluorescence quantitative PCR can be seen in the original hematopoietic transcription factor lmo2 and scl,the experimental group from 9 hpf to 24 hpf significantly lower than the other two groups.In the targeted hematopoietic transcription factor,c-myb experimental group from 12 hpf to 24 hpf significantly lower than the other two groups,runx1 experimental group from 9 hpf to 30 hpf significantly lower than the other two groups.Myeloid hematopoietic transcription factor pu.1 and erythroid hematopoietic transcription factor gata-1,the experimental group from 9 hpf to 30 hpf were significantly lower than the other two groups.The differences were statistically significant.Conclusion Irx5a gene plays an inhibitory role in the transcriptional regulators of primitive hematopoiesis,adult hematopoiesis,erythroid hematopoiesis and myeloid hematopoiesis in the zebrafish.%目的 探讨易洛魁族同源盒5a(Irx5a)基因对斑马鱼胚胎早期造血相关转录调控因子的影响.方法 于斑马鱼胚胎1细胞受精卵期,显微注射Irx5a-EGFP mRNA斑马鱼胚胎作为实验组、显微注射增强型绿色荧光蛋白(EGFP) mRNA斑马鱼胚胎作为实验对照组、野生型斑马鱼作为空白对照组,应用qPCR检测相关造血转录调控因子在斑马鱼胚胎受精后9 h(9 hpf)、12h、18h、24 h和30 h时相的表达水平.结果 实时荧光定量PCR结果表明,在原始造血转录因子lmo2、scl中,实验组从9 hpf～24 hpf较另外两组间明显减低;在定向造血转录因子中,c-myb实验组从12 hpf～24 hpf较另外两组间明显减低,runx1实验组从9 hpf～30 hpf较另外两组间明显减低;髓系造血转录因子pu.1和红系造血转录调控因子gata-1实验组从9 hpf～30 hpf较另外两组间明显减低,且差异有统计学意义.结论 Irx5a基因在斑马鱼原始造血、成体造血、红系造血、髓系造血的重要转录调控因子中发挥着抑制作用.