Zfp36l1b protects angiogenesis through Notch1b/Dll4 and Vegfa regulation in zebrafish
Zfp36l1b protects angiogenesis through Notch1b/Dll4 and Vegfa regulation in zebrafish
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Zfp36l1b 通过 Notch1b/Dll4 和 Vegfa 调节斑马鱼保护血管生成
DOI：
10.1016/j.atherosclerosis.2020.07.021
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发表时间：
2020-09-01
期刊：
影响因子：
5.3
通讯作者：
Jing, Qing
中科院分区：
文献类型：
--
作者：
Hu, Yang-Xi;Zhu, Rong-Fang;Jing, Qing
背景与目的：血管生成是胚胎发生过程中血管功能建立的关键过程，涉及多种信号机制。据报道，RNA结合蛋白Zfp36l1参与了不同物种的各种疾病，包括心血管疾病。然而，Zfp36l1b作为Zfp36l1在斑马鱼中的2个类似物之一，是否与哺乳动物的Zfp36l1一样工作，以及是否存在不同的分子机制尚不清楚。在这里，我们发现Zfp36l1b在斑马鱼胚胎血管生成中起着至关重要的保护作用。方法：采用透明转基因斑马鱼和野生型斑马鱼幼体，在体内共聚焦实验中，对Zfp36l1b基因被morpholinos （MOs）敲低后的早期血管生成过程进行动态观察。采用原位杂交和荧光活化细胞分选检测Zfp36l1b的表达。通过mRNA拯救、CRISPR/Cas9敲低、荧光素酶报告基因实验进一步探讨Zfp36l1b在血管生成中的作用。结果：我们发现，敲低Zfp36l1b可导致胞间血管和肠下静脉（SIVs）血管生成缺陷，而添加Zfp36l1b mRNA可挽救SIVs。此外，敲低Zfp36l1b可抑制Notch1b的表达，而敲低Notch1b可部分缓解Zfp36l1b下调诱导的血管生成缺陷。此外，Zfp36l1b敲低可减轻Vegfa过表达引起的siv过度分支。结论：我们的研究结果表明Zfp36l1b通过抑制Notch1b/Fll4及与Vegfa的协同作用，部分通过影响内皮尖细胞丝状伪足的延伸和内皮细胞的增殖，从而参与正常血管回路的建立。
Background and aims: Angiogenesis is a key process for establishing functional vasculature during embryogenesis and involves different signaling mechanisms. The RNA binding protein Zfp36l1 was reported to be involved in various diseases in different species, including cardiovascular diseases. However, whether Zfp36l1b, one of the 2 paralogs of Zfp36l1 in zebrafish, works like mammalian Zfp36l1, and if the molecular mechanisms are different remains unclear. Here, we show that Zfp36l1b plays a crucial protective role in angiogenesis of zebrafish embryos.Methods: We used transparent transgenic and wild-type zebrafish larvae to dynamically investigate the early stage of angiogenesis with confocal in vivo, after the knockdown of Zfp36l1b by morpholinos (MOs). In situ hybridization and fluorescence-activated cell sorting were performed to detect Zfp36l1b expression. mRNA rescue and CRISPR/Cas9 knockdown, and luciferase reporter experiments were performed to further explore the role of Zfp36l1b in angiogenesis.Results: We found that knockdown of Zfp36l1b led to defected angiogenesis in intersomitic vessels and subintestinal veins (SIVs), which could be rescued by the addition of Zfp36l1b mRNA. Moreover, knockdown of Zfp36l1b suppressed Notch1b expression, while knockdown of Notch1b resulted in a partial relief of angiogenesis defects induced by Zfp36l1b down-regulation. Besides, Zfp36l1b knockdown alleviated the excessive branch of SIVs caused by Vegfa over-expression.Conclusions: Our results show that Zfp36l1b is responsible for establishing normal vessel circuits by affecting the extension of endothelial tip cells filopodia and the proliferation of endothelial cells partly through Notch1b/Fll4 suppression and synergistic function with Vegfa.