Chen Kan, Fan Yuqi, Gu Jun, Han Zhihua, Wang Yue, Gao Lin, Zeng Huasu, Mao Chengyu, Wang Changqian
Department of Cardiology, Shanghai Ninth People's Hospital Affiliated Shanghai Jiaotong University School of Medicine, No. 639 Zhizaoju Road, Shanghai, 200011, China.
Transgenic Res. 2021 Dec;30(6):739-750. doi: 10.1007/s11248-021-00276-5. Epub 2021 Aug 4.
Our study was aimed to investigate the effects of lgals3a (Gal-3 encoding gene) on the development of zebrafish embryo and its underlying mechanisms. Morpholino (MO) technology was used to inhibit the expression of zebrafish lgals3a, and the effect of lgals3a gene knockdown on zebrafish embryo development and the number of monocyte macrophages was observed. Effect of lgals3a-e3i3-MO on apoptosis of zebrafish was detected by acridine orange staining. In addition, the mRNA expression levels of Wnt/β-catenin signaling pathway-related genes were detected by RT-qPCR. Compared with control-MO group, the zebrafish embryos injected with lgals3a-e3i3-MO had obvious defects in the head, eyes, and tail, and pericardial edema. Lgals3a-e3i3-MO significantly reduced the number of mononuclear macrophages in zebrafish embryos compared with the control-MO group. The results of acridine orange staining showed that compared with the control-MO group, lgals3a-e3i3-MO promoted cardiomyocyte apoptosis in zebrafish. Furthermore, lgals3a-e3i3-MO significantly up-regulated the expression of dkk1b, wnt9a, lrp5, fzd7a, β-catenin, Gsk-3β, mycn, myca in the Wnt/β-catenin pathway, and decreased the expression of lef1. These results indicate that lgals3a-e3i3-MO inhibits zebrafish embryo development, reduces the number of mononuclear macrophages, activates Wnt/β-catenin signaling pathway and promotes cardiomyocyte apoptosis.
我们的研究旨在探讨lgals3a(半乳糖凝集素-3编码基因)对斑马鱼胚胎发育的影响及其潜在机制。使用吗啉代寡核苷酸(MO)技术抑制斑马鱼lgals3a的表达,并观察lgals3a基因敲低对斑马鱼胚胎发育和单核巨噬细胞数量的影响。通过吖啶橙染色检测lgals3a-e3i3-MO对斑马鱼凋亡的影响。此外,通过逆转录定量聚合酶链反应(RT-qPCR)检测Wnt/β-连环蛋白信号通路相关基因的mRNA表达水平。与对照MO组相比,注射lgals3a-e3i3-MO的斑马鱼胚胎在头部、眼睛和尾部出现明显缺陷,并伴有心包水肿。与对照MO组相比,lgals3a-e3i3-MO显著减少了斑马鱼胚胎中的单核巨噬细胞数量。吖啶橙染色结果显示,与对照MO组相比,lgals3a-e3i3-MO促进了斑马鱼心肌细胞凋亡。此外,lgals3a-e3i3-MO显著上调了Wnt/β-连环蛋白通路中dkk1b、wnt9a、lrp5、fzd7a、β-连环蛋白、Gsk-3β、mycn、myca的表达,并降低了lef1的表达。这些结果表明,lgals3a-e3i3-MO抑制斑马鱼胚胎发育,减少单核巨噬细胞数量,激活Wnt/β-连环蛋白信号通路并促进心肌细胞凋亡。