Irx5a基因过表达对斑马鱼胚胎早期造血的影响

doi:10.3969/j.issn.1674-5817.2017.04.011

实验动物与比较医学 ›› 2017, Vol. 37 ›› Issue (4): 309-314.DOI: 10.3969/j.issn.1674-5817.2017.04.011

• 第十六届中国西部实验动物管理与学术研讨会论文专题 • 上一篇下一篇

Irx5a基因过表达对斑马鱼胚胎早期造血的影响

金璐^1,2, 李燕², 李志操^1,2, 吴西军¹, 周艳华¹, 何志旭^1,2, 舒莉萍^1,3,4

贵州医科大学 1.组织工程和干细胞实验中心; 2.儿童医学中心; 3.基础医学院免疫学教研室; 4.实验动物中心, 贵阳550004

收稿日期:2017-05-30 出版日期:2017-08-25 发布日期:2017-08-25
作者简介:金璐(1991-),女,临床医学硕士研究生,贵州医科大学组织工程和干细胞实验中心,E-mail:326687520@qq.com
基金资助:
国家自然科学基金(31460312)

Effects of Irx5a Gene Overexpression on Early Hematopoietic in Zebrafish Embryos

JIN Lu^1,2, LI Yan², LI Zhi-cao^1,2, WU Xi-jun¹, ZHOU Yan-hua¹, HE Zhi-xu^1,2, SHU Li-ping^1,3,4

1. Tissue Engineering and Stem Cell Research Center, 2. Deptartment of Pediatrics, 3. Department of Immunology, 4. Laboratory Animal Center,Guizhou Medical University, Guiyang 550004, China

Received:2017-05-30 Online:2017-08-25 Published:2017-08-25

摘要/Abstract

摘要： 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)、12 h、18 h、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。

关键词: 斑马鱼, 5a(Irx5a), 造血转录调控因子

Abstract: 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 transcription-PCR (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.

Key words: Zebrafish, Iroquois homebox 5a(Irx5a), Hematopoietic transcription factor

中图分类号:

Q95-33

金璐, 李燕, 李志操, 吴西军, 周艳华, 何志旭, 舒莉萍. Irx5a基因过表达对斑马鱼胚胎早期造血的影响[J]. 实验动物与比较医学, 2017, 37(4): 309-314.

JIN Lu, LI Yan, LI Zhi-cao, WU Xi-jun, ZHOU Yan-hua, HE Zhi-xu, SHU Li-ping. Effects of Irx5a Gene Overexpression on Early Hematopoietic in Zebrafish Embryos[J]. Laboratory Animal and Comparative Medicine, 2017, 37(4): 309-314.

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Irx5a基因过表达对斑马鱼胚胎早期造血的影响

Effects of Irx5a Gene Overexpression on Early Hematopoietic in Zebrafish Embryos

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