武汉轻工大学学报2024,Vol.43Issue(2) :50-55.DOI:10.3969/j.issn.2095-7386.2024.02.007

Hif-1α在斑马鱼吞噬细胞迁移及发育中的生物学功能

Biological function of Hif-1α in phagocyte migration and development of zebrafish

汪豪 罗舸洋 魏泽昊 司马重圆 晏博 张舒林 余晓丽
武汉轻工大学学报2024,Vol.43Issue(2) :50-55.DOI:10.3969/j.issn.2095-7386.2024.02.007

Hif-1α在斑马鱼吞噬细胞迁移及发育中的生物学功能

Biological function of Hif-1α in phagocyte migration and development of zebrafish

汪豪 1罗舸洋 2魏泽昊 1司马重圆 1晏博 2张舒林 1余晓丽1
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作者信息

  • 1. 武汉轻工大学生命科学与技术学院,武汉 430023
  • 2. 上海市公共卫生临床中心,上海 201508
  • 折叠

摘要

探究斑马鱼低氧诱导因子-1α(Hif-1α)基因在中性粒细胞和巨噬细胞的发育和迁移中所起的作用.将斑马鱼分为野生对照组和Hif-1α基因突变组.使用CRISPR/Cas9技术对斑马鱼的Hif-1α基因进行突变.随后,通过苏丹黑B染色法观察了野生对照组和Hif-1α基因突变组中性粒细胞的染色情况,以及通过中性红染色法观察了巨噬细胞的染色情况并对结果进行了统计分析.结果显示成功构建缺失8个碱基的斑马鱼Hif-1α基因突变体,且Hif-1α基因突变的斑马鱼幼鱼发育3 d后,中性粒细胞迁移到急性损伤部位的数量显著减少,突变型斑马鱼后脑室中的小胶质巨噬细胞数量比野生型显著增多.综上,本研究成功构建了斑马鱼Hif-1α基因纯合突变体,同时表明Hif-1α基因具有抑制中性粒细胞向炎症区域迁移的作用,并促进脑室巨噬细胞的生长发育,为后续研究急性损伤模型提供基础.

Abstract

Investigating the role of the zebrafish hypoxia-inducible factor-1 alpha(Hif-1α)gene in the devel-opment and migration of neutrophils and macrophages.Zebrafish were divided into wild-type control group and Hif-1α gene mutant group.CRISPR/Cas9 technology was utilized to mutate the Hif-1α gene in ze-brafish.Subsequently,Sudan Black B staining was performed to observe the staining of neutrophils in the wild-type control group and Hif-1α gene mutant group,and Neutral Red staining was employed to observe the staining of macrophages,followed by statistical analysis of the results.The results demonstrated suc-cessfully construction of zebrafish Hif-1α gene mutant with deletion of 8 bases.Furthermore,after 3 days of development in zebrafish larvae with mutated Hif-1α gene,there was a significant decrease in the number of neutrophils migrating to acute injury sites,while the number of microglial macrophages in the posterior ventricle of mutant zebrafish significantly increased compared to the wild-type.In summary,this study suc-cessfully constructed a zebrafish Hif-1α gene homozygous mutant.Additionally,it elucidated the role of Hif-1α gene in inhibiting neutrophil migration to inflammatory regions and promoting the growth and de-velopment of ventricular macrophages,laying a foundation for subsequent studies on acute injury models.

关键词

Hif-1α基因纯合突变体/斑马鱼幼鱼/中性粒细胞迁移/巨噬细胞发育

Key words

Hif-1α gene homozygous mutants/zebrafish larvae/neutrophil migration/macrophage develop-ment

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基金项目

国家重点研发国际合作项目(2023YFE0199200)

武汉市科技局2020年度应用基础前沿项目(2020020601012231)

出版年

2024
武汉轻工大学学报
武汉工业学院

武汉轻工大学学报

影响因子:0.356
ISSN:1009-4881
参考文献量16
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