A zebrafish screen for craniofacial mutants identifies wdr68 as a highly conserved gene required for endothelin-1 expression.
A zebrafish screen for craniofacial mutants identifies wdr68 as a highly conserved gene required for endothelin-1 expression.
复制标题
颅面突变体的斑马鱼筛选将WDR68识别为内皮素-1表达所需的高度保守基因。
DOI：
10.1186/1471-213x-6-28
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发表时间：
2006-06-07
影响因子：
--
通讯作者：
Hopkins N
中科院分区：
文献类型：
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作者：
Nissen RM;Amsterdam A;Hopkins N
颅面出生缺陷是颅神经嵴（NC）形态和形态发生缺陷的结果。脊椎动物颅面骨骼来源于颅NC细胞，这些细胞的图案发生在咽弓内。大量的努力导致了颅面骨骼发育所需的几个基因的鉴定，如内皮素-1 （edn1）信号通路，这是下颌形成所必需的。然而，颅面发育所需的许多必要基因仍有待确定。通过筛选含有约25%胚胎发育必需基因的插入性斑马鱼突变体，我们鉴定出颅面发育所需的15个必需基因。我们确定了下颌发育所需的3个基因。我们还鉴定了斑马鱼的camomelic Dysplasia和Ehlers-Danlos综合征模型。为了进一步证明该方法的实用性，我们对wdr68基因进行了表征。我们发现，wdr68作用于edn1通路的上游，也是形成上颌等级物腭合体所必需的。我们还提供证据表明，edn1通路功能所需的wdr68活性水平在第一和第二弓之间是不同的。Wdr68分别与胚胎生长和肌管分化所需的两种微型脑相关激酶Dyrk1a和Dyrk1b相互作用。我们发现GFP-Wdr68融合蛋白与Dyrk1a定位于细胞核，而与之相反的是，工程功能缺失突变wdr68 - t284f不再积聚在细胞核中，无法挽救wdr68突变动物。Wdr68同源物似乎存在于所有真核生物基因组中。值得注意的是，我们发现即使昆虫缺乏NC细胞谱系，果蝇wdr68同源物CG14614也可以替代脊椎动物的wdr68基因。这项工作代表了对颅面发育所需的大约25%的必要基因的系统鉴定。两种人类疾病综合征的斑马鱼模型的鉴定表明，与其他基因的同源物也可能与人类颅面发育有关。wdr68的初步表征表明高度保守的wdr68 - dyrk1蛋白复合物在颅面发育中起重要作用。
Craniofacial birth defects result from defects in cranial neural crest (NC) patterning and morphogenesis. The vertebrate craniofacial skeleton is derived from cranial NC cells and the patterning of these cells occurs within the pharyngeal arches. Substantial efforts have led to the identification of several genes required for craniofacial skeletal development such as the endothelin-1 (edn1) signaling pathway that is required for lower jaw formation. However, many essential genes required for craniofacial development remain to be identified. Through screening a collection of insertional zebrafish mutants containing approximately 25% of the genes essential for embryonic development, we present the identification of 15 essential genes that are required for craniofacial development. We identified 3 genes required for hyomandibular development. We also identified zebrafish models for Campomelic Dysplasia and Ehlers-Danlos syndrome. To further demonstrate the utility of this method, we include a characterization of the wdr68 gene. We show that wdr68 acts upstream of the edn1 pathway and is also required for formation of the upper jaw equivalent, the palatoquadrate. We also present evidence that the level of wdr68 activity required for edn1 pathway function differs between the 1st and 2nd arches. Wdr68 interacts with two minibrain-related kinases, Dyrk1a and Dyrk1b, required for embryonic growth and myotube differentiation, respectively. We show that a GFP-Wdr68 fusion protein localizes to the nucleus with Dyrk1a in contrast to an engineered loss of function mutation Wdr68-T284F that no longer accumulated in the cell nucleus and failed to rescue wdr68 mutant animals. Wdr68 homologs appear to exist in all eukaryotic genomes. Notably, we found that the Drosophila wdr68 homolog CG14614 could substitute for the vertebrate wdr68 gene even though insects lack the NC cell lineage. This work represents a systematic identification of approximately 25% of the essential genes required for craniofacial development. The identification of zebrafish models for two human disease syndromes indicates that homologs to the other genes are likely to also be relevant for human craniofacial development. The initial characterization of wdr68 suggests an important role in craniofacial development for the highly conserved Wdr68-Dyrk1 protein complexes.