Microtubule Severing Protein Fignl2 Contributes to Endothelial and Neuronal Branching in Zebrafish Development.
Microtubule Severing Protein Fignl2 Contributes to Endothelial and Neuronal Branching in Zebrafish Development.
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微管切断蛋白 Fignl2 有助于斑马鱼发育中的内皮和神经元分支
DOI：
10.3389/fcell.2020.593234
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发表时间：
2020
影响因子：
5.5
通讯作者：
Liu M
中科院分区：
文献类型：
--
作者：
Dong Z;Chen X;Li Y;Zhuo R;Lai X;Liu M
此前，fidgetin（fign）及其家族成员fidgetin-like 1（fignl 1）和fidgetin-like 2（fignl 2）被发现在斑马鱼脑发育过程中高度表达，提示其在神经系统中的功能。在这项研究中，我们报告了这些基因功能丧失对发育的影响。我们设计并鉴定了靶向产生fign、fignl 1和fignl 2突变体的单向导RNA，然后观察了整体形态和行为变化。我们的研究结果表明，虽然fign和fignl 1无效突变体显示没有显着的缺陷，fignl 2无效斑马鱼突变体显示心包水肿，心率降低，眼睛变小; fignl 2无效突变体响应的光暗转换与游泳速度较低。图12通过原位杂交和重新分析单细胞RNAseq结果的在线数据集，在血管内皮细胞中鉴定mRNA。最后，我们使用吗啉代寡核苷酸证实fignl 2敲低导致严重的心脏水肿，这是由异常的血管分支引起的。斑马鱼的fignl 2变形体也表现出较长的轴突长度和更多的分支的尾部初级神经元。综上所述，我们总结了Fignl 2在内皮细胞和神经元中的细胞分支上的功能。这项研究首次报道了微管切断蛋白Fignl 2在发育过程中有助于细胞分支。
Previously, fidgetin (fign) and its family members fidgetin-like 1 (fignl1) and fidgetin-like 2 (fignl2) were found to be highly expressed during zebrafish brain development, suggesting their functions in the nervous system. In this study, we report the effects of loss-of-function of these genes on development. We designed and identified single-guide RNAs targeted to generate fign, fignl1, and fignl2 mutants and then observed the overall morphological and behavioral changes. Our findings showed that while fign and fignl1 null mutants displayed no significant defects, fignl2 null zebrafish mutants displayed pericardial edema, reduced heart rate, and smaller eyes; fignl2 null mutants responded to the light-darkness shift with a lower swimming velocity. fignl2 mRNAs were identified in vascular endothelial cells by in situ hybridization and re-analysis of an online dataset of single-cell RNAseq results. Finally, we used morpholino oligonucleotides to confirm that fignl2 knockdown resulted in severe heart edema, which was caused by abnormal vascular branching. The zebrafish fignl2 morphants also showed longer axonal length and more branches of caudal primary neurons. Taken together, we summarize that Fignl2 functions on cellular branches in endothelial cells and neurons. This study reported for the first time that the microtubule-severing protein Fignl2 contributes to cell branching during development.