Department of Haematology, Centre for Cancer Biology, SA Pathology, Frome Rd, Adelaide, SA 5000, Australia.
Cell Death Differ. 2010 Aug;17(8):1302-14. doi: 10.1038/cdd.2010.12. Epub 2010 Feb 12.
The centrosome is the primary microtubule organising centre of the cell. It is composed of many proteins, some of which make up the core of the centrosome, whereas others are used for specific functions. Although the cellular roles of many centrosomal proteins are well defined, much less is known about their functions and the role of the centrosome in development. In this study we investigated the function of NEDD1, a critical component of the centrosome essential for microtubule nucleation, in zebrafish (Danio rerio) development. The zebrafish homologue of NEDD1 (zNEDD1) was cloned and found to have a similar localisation and function to mammalian NEDD1. We show that zNEDD1 is essential for survival, as a high level of knockdown was embryonic lethal. Partial knockdown of zNEDD1 caused abnormalities including an increase in mitotic and apoptotic cells. Pronounced phenotypic defects were seen in the brain, with a lack of defined brain structures, incomplete neural tube formation and a disorganisation of neurons. In addition, we show that a reduction in zNEDD1 resulted in the loss of gamma-tubulin at the centrosome. Our data thus demonstrate that zNEDD1 is critical for the recruitment of gamma-tubulin to the centrosome, and is essential for the proper development of zebrafish.
中心体是细胞的主要微管组织中心。它由许多蛋白质组成,其中一些构成了中心体的核心,而另一些则用于特定的功能。尽管许多中心体蛋白的细胞功能已经得到很好的定义,但人们对它们的功能以及中心体在发育中的作用知之甚少。在这项研究中,我们研究了 NEDD1 在斑马鱼(Danio rerio)发育中的作用,NEDD1 是微管成核所必需的中心体的关键组成部分。克隆了斑马鱼 NEDD1 的同源物(zNEDD1),并发现其与哺乳动物 NEDD1 的定位和功能相似。我们表明 zNEDD1 对生存至关重要,因为高水平的敲低导致胚胎致死。zNEDD1 的部分敲低导致包括有丝分裂和细胞凋亡细胞增加的异常。在大脑中观察到明显的表型缺陷,缺乏明确的脑结构,神经管形成不完全,神经元排列紊乱。此外,我们还表明,zNEDD1 的减少导致中心体中γ-微管蛋白的丢失。因此,我们的数据表明,zNEDD1 对于招募γ-微管蛋白到中心体至关重要,并且对于斑马鱼的正常发育是必不可少的。