Mazmanian Gohar, Kovshilovsky Michael, Yen Debbie, Mohanty Aditya, Mohanty Sudipta, Nee Alex, Nissen Robert M
Department of Biological Sciences, California State University Los Angeles, 5151 State University Drive, Los Angeles, CA 90032, USA.
Genesis. 2010 Jan;48(1):20-30. doi: 10.1002/dvg.20578.
Nodal-signaling is required for specification of mesoderm, endoderm, establishing left-right asymmetry, and craniofacial development. Wdr68 is a WD40-repeat domain-containing protein recently shown to be required for endothelin-1 (edn1) expression and subsequent lower jaw development. Previous reports detected the Wdr68 protein in multiprotein complexes containing mammalian members of the dual-specificity tyrosine-regulated kinase (dyrk) family. Here we describe the characterization of the zebrafish dyrk1b homolog. We report the detection of a physical interaction between Dyrk1b and Wdr68. We also found perturbations of nodal signaling in dyrk1b antisense morpholino knockdown (dyrk1b-MO) animals. Specifically, we found reduced expression of lft1 and lft2 (lft1/2) during gastrulation and a near complete loss of the later asymmetric lft1/2 expression domains. Although wdr68-MO animals did not display lft1/2 expression defects during gastrulation, they displayed a near complete loss of the later asymmetric lft1/2 expression domains. While expression of ndr1 was not substantially effected during gastrulation, ndr2 expression was moderately reduced in dyrk1b-MO animals. Analysis of additional downstream components of the nodal signaling pathway in dyrk1b-MO animals revealed modestly expanded expression of the dorsal axial mesoderm marker gsc while the pan-mesodermal marker bik was largely unaffected. The endodermal markers cas and sox17 were also moderately reduced in dyrk1b-MO animals. Notably, and similar to defects previously reported for wdr68 mutant animals, we also found reduced expression of the pharyngeal pouch marker edn1 in dyrk1b-MO animals. Taken together, these data reveal a role for dyrk1b in endoderm formation and craniofacial patterning in the zebrafish.
中胚层、内胚层的特化,左右不对称性的建立以及颅面发育都需要节点信号传导。Wdr68是一种含有WD40重复结构域的蛋白质,最近被证明是内皮素-1(edn1)表达及随后的下颌发育所必需的。先前的报道在包含双特异性酪氨酸调节激酶(dyrk)家族哺乳动物成员的多蛋白复合物中检测到了Wdr68蛋白。在此,我们描述斑马鱼dyrk1b同源物的特征。我们报告了Dyrk1b与Wdr68之间存在物理相互作用。我们还发现,在dyrk1b反义吗啉代敲低(dyrk1b-MO)动物中,节点信号传导受到干扰。具体而言,我们发现在原肠胚形成期间,lft1和lft2(lft1/2)的表达降低,并且后期不对称的lft1/2表达域几乎完全丧失。虽然wdr68-MO动物在原肠胚形成期间未表现出lft1/2表达缺陷,但它们后期不对称的lft1/2表达域几乎完全丧失。虽然在原肠胚形成期间ndr1的表达没有受到实质性影响,但在dyrk1b-MO动物中ndr2的表达适度降低。对dyrk1b-MO动物中节点信号通路的其他下游成分进行分析发现,背侧轴向中胚层标记物gsc的表达适度扩大,而泛中胚层标记物bik基本未受影响。内胚层标记物cas和sox17在dyrk1b-MO动物中也适度降低。值得注意的是,与先前报道的wdr68突变动物的缺陷类似,我们还发现dyrk1b-MO动物中咽囊标记物edn1的表达降低。综上所述,这些数据揭示了dyrk1b在斑马鱼内胚层形成和颅面模式形成中的作用。