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

作　　者：汪豪 罗舸洋 魏泽昊 司马重圆 晏博 张舒林 余晓丽[1] WANG Hao;LUO Geyang;WEI Zehao;SIMA Chongyuan;YAN Bo;ZHANG Shulin;YU Xiaoli(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China;Shanghai Public Health Clinical Center,Shanghai 201508,China)

机构地区：[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

出　　处：《武汉轻工大学学报》2024年第2期50-55,共6页Journal of Wuhan Polytechnic University

基　　金：国家重点研发国际合作项目(编号:2023YFE0199200);武汉市科技局2020年度应用基础前沿项目(编号:2020020601012231).

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。Investigating the role of the zebrafish hypoxia-inducible factor-1 alpha(Hif-1α)gene in the development and migration of neutrophils and macrophages.Zebrafish were divided into wild-type control group and Hif-1αgene mutant group.CRISPR/Cas9 technology was utilized to mutate the Hif-1αgene in zebrafish.Subsequently,Sudan Black B staining was performed to observe the staining of neutrophils in the wild-type control group and Hif-1αgene mutant group,and Neutral Red staining was employed to observe the staining of macrophages,followed by statistical analysis of the results.The results demonstrated successfully construction of zebrafish Hif-1αgene mutant with deletion of 8 bases.Furthermore,after 3 days of development in zebrafish larvae with mutated Hif-1αgene,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 successfully constructed a zebrafish Hif-1αgene homozygous mutant.Additionally,it elucidated the role of Hif-1αgene in inhibiting neutrophil migration to inflammatory regions and promoting the growth and development 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 

分 类 号：Q344.1[生物学—遗传学]