A globin-family protein, Cytoglobin 1, is involved in the development of neural crest-derived tissues and organs in zebrafish
A globin-family protein, Cytoglobin 1, is involved in the development of neural crest-derived tissues and organs in zebrafish
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珠蛋白家族蛋白细胞珠蛋白 1 参与斑马鱼神经嵴衍生组织和器官的发育
DOI：
10.1016/j.ydbio.2020.12.016
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发表时间：
2021
影响因子：
2.7
通讯作者：
Yamasu Kyo
中科院分区：
文献类型：
--
作者：
Takahashi Kazuki;Ito Yuki;Yoshimura Mami;Nikaido Masataka;Yuikawa Tatsuya;Kawamura Akinori;Tsuda Sachiko;Kage Daichi;Yamasu Kyo
斑马鱼是一种很好的模式动物，适合正向遗传学方法。为了揭示脊椎动物未知的发育调控机制，我们进行了化学诱变筛选，并确定了一个新的突变，kanazutsi（kzt）。这种突变是隐性的，其纯合子是胚胎致死的。突变胚胎遭受了各种形态缺陷，如头部扁平化，心包水肿，循环缺陷，黑色素细胞分布模式中断，侏儒眼睛，有缺陷的下巴，以及头部的广泛凋亡，这表明主要受影响的组织来自神经嵴细胞（NCC）。组织特异性标记物inkzt突变体的表达表明，NCC的早期特化是正常的，但其后期分化受到严重影响。该突变被映射到3号染色体的连锁分析，nearcytoglobin 1（cygb 1），其产品是一个球蛋白家族的呼吸蛋白。cygb 1的表达被激活在体节和颅NCC在野生型胚胎的体节形成过程中，但显着下调突变胚胎，尽管正常的基因产物的一级结构。kzt突变被低剂量morpholino oligos敲低cygb 1表型复制，并被cygb 1过表达部分挽救。通过CRISPR/Cas9技术建立的cygb 1的严重敲除和无效突变都在早期阶段导致了更严重的缺陷。因此，很可能cygb 1的下调导致了kzt突变的许多（如果不是全部的话）表型。这些结果揭示了脊椎动物胚胎中对珠蛋白家族蛋白的需求，特别是在NCC的分化和随后的发育中。
The zebrafish is an excellent model animal that is amenable to forward genetics approaches. To uncover unknown developmental regulatory mechanisms in vertebrates, we conducted chemical mutagenesis screening and identified a novel mutation,kanazutsi(kzt). This mutation is recessive, and its homozygotes are embryonic lethal. Mutant embryos suffered from a variety of morphological defects, such as head flattening, pericardial edema, circulation defects, disrupted patterns of melanophore distribution, dwarf eyes, a defective jaw, and extensive apoptosis in the head, which indicates that the main affected tissues are derived from neural crest cells (NCCs). The expression of tissue-specific markers inkztmutants showed that the early specification of NCCs was normal, but their later differentiation was severely affected. The mutation was mapped to chromosome 3 by linkage analyses, nearcytoglobin 1(cygb1), the product of which is a globin-family respiratory protein.cygb1expression was activated during somitogenesis in somites and cranial NCCs in wild-type embryos but was significantly downregulated in mutant embryos, despite the normal primary structure of the gene product. Thekztmutation was phenocopied bycygb1knockdown with low-dose morpholino oligos and was partially rescued bycygb1overexpression. Both severe knockdown and null mutation ofcygb1, established by the CRISPR/Cas9 technique, resulted in far more severe defects at early stages. Thus, it is highly likely that the downregulation ofcygb1is responsible for many, if not all, of the phenotypes of thekztmutation. These results reveal a requirement for globin family proteins in vertebrate embryos, particularly in the differentiation and subsequent development of NCCs.