Nusap1 is essential for neural crest cell migration in zebrafish
Nusap1 is essential for neural crest cell migration in zebrafish
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Nusap1 对于斑马鱼神经嵴细胞迁移至关重要
DOI：
10.1007/s13238-010-0036-8
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发表时间：
2010-03-01
期刊：
影响因子：
21.1
通讯作者：
Huang, Huizhe
中科院分区：
文献类型：
--
作者：
Nie, Jing;Wang, Hua;Huang, Huizhe
微管在有丝分裂纺锤体组装和染色体分离中起重要作用，维持细胞周期的正常进行。在上皮细胞和神经细胞培养物中已经鉴定出许多微管相关蛋白，然而，由于缺乏适当的体内动物模型，它们的生理意义还没有得到很好的表征。核仁纺锤体相关蛋白（NuSAP）是一种微管结合蛋白，细胞培养研究发现其参与有丝分裂。在这份报告中，我们确定了斑马鱼同源的人类NuSAP和研究其表达谱和功能。利用原位杂交技术，我们证明了斑马鱼nusap1的转录本在视网膜、前脑、后脑和神经嵴中特异性表达。当nusap1在体内的表达通过反义寡核苷酸morpholino技术敲低，nusap1的morphants显示受损的形态发生在躯干和蛋黄延伸，这意味着Nusap1参与细胞迁移。机制研究表明，nusap1吗啡肽改变了神经嵴标记物scretin和sox 9b的表达模式，但血管和脊索标记物sgata1和shh的表达正常。此外，nusap1 mRNA注射导致视网膜和后脑组织严重的凋亡，这些表型可以通过共注射morpholino againstnusap1来挽救。这些观察结果不仅表明Nusap1在连接细胞凋亡与细胞迁移中的作用，而且还提供了强有力的证据表明Nusap1可能参与脊椎动物的形态发生。
Microtubules play important roles in mitotic spindle assembly and chromosome segregation to maintain normal cell cycle progression. A number of microtubule-associated proteins have been identified in epithelial and neural cell cultures; however, their physiological significance is not well characterized due to the lack of appropriatein vivoanimal models. Nucleolar spindle-associated protein (NuSAP) is a microtubule-binding protein and is reported to be involved in mitosis by cell culture studies. In this report, we identified the zebrafish homologue of human NuSAP and investigated its expression profile and functions. Usingin situhybridization, we demonstrated that transcripts of zebrafishnusap1are specifically expressed in the retina, forebrain, hindbrain and neural crest. When thein vivoexpression ofnusap1was knocked down through antisense oligonucleotide morpholino technology, the morphants ofnusap1showed impaired morphogenesis in the trunk and yolk extension, implying the involvement of Nusap1 in cell migration. Mechanistic studies revealed thatnusap1morphants have an altered expression pattern of neural crest markerscrestinandsox9b, but normal expression of blood vessel and notochord markersgata1andshh. In addition,nusap1mRNA injection caused serious apoptosis in retina and hindbrain tissue, and these phenotypes can be rescued by co-injection of morpholino againstnusap1. These observations not only suggest a role for Nusap1 in connecting apoptosis with cell migration, but also provide strong evidences that Nusap1 is potentially involved in morphogenesis in vertebrates.