Wnt signaling interacts with bmp and edn1 to regulate dorsal-ventral patterning and growth of the craniofacial skeleton.
Wnt signaling interacts with bmp and edn1 to regulate dorsal-ventral patterning and growth of the craniofacial skeleton.
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DOI：
10.1371/journal.pgen.1004479
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发表时间：
2014-07
期刊：
影响因子：
4.5
通讯作者：
Schilling TF
中科院分区：
文献类型：
--
作者：
Alexander C;Piloto S;Le Pabic P;Schilling TF
颅面的发育需要来自上皮细胞的信号来形成骨骼发生神经嵴（NC）细胞，例如将每个咽弓细分为不同的背侧（D）和腹侧（V）元件。Wnt信号转导在NC和颅面发育的许多方面都有牵连，但其在D-V弓模式中的作用仍不清楚。为了解决这个问题，我们以时间控制的方式阻断了斑马鱼胚胎中的Wnt信号传导，在NC迁移后使用转基因过表达显性负性Tcf 3（dntcf 3），（Tg（hsp 70 i：tcf 3-GFP））或经典Wnt抑制剂dickkopf 1（dkk 1），（Tg（hsp 70 i：dkk 1-GFP）。在dntcf 3转基因中，热休克胚胎的腹弓中的NC细胞显示出增殖减少、腹侧图案基因（hand 2、dlx 3b、dlx 5a、msxe）的表达和腹侧软骨分化（例如下颌）。这些D-V图案缺陷类似于缺乏Bmp或Edn 1信号传导的斑马鱼胚胎的表型，并且dntcf 3的过表达显著降低了弓中Bmp受体的子集的表达。异位BMP（或EDN 1）蛋白的添加部分挽救腹侧发育和表达的dlx 3b，dlx 5a，和msxe在Wnt信号缺陷的胚胎，但令人惊讶的是，不抢救手2的表达。因此，Wnt信号提供了腹型图案线索拱NC细胞，部分通过调节BMP和Edn 1信号，但独立调节手2。类似地，热休克dkk 1+胚胎表现出腹弓减少，但也有下颌裂在腹中线没有看到dntcf 3+胚胎。Dkk 1在咽内胚层中表达，细胞移植实验表明，dntcf 3必须在咽内胚层中过表达，以破坏D-V弓模式，这表明Wnt和Wnt拮抗剂的不同内胚层作用模式发育中的骨骼。颅面畸形是最常见的出生缺陷之一。了解颅面疾病的分子机制对于制定治疗策略至关重要。颅面骨骼的大部分是由称为咽弓的特殊胚胎结构形成的。这些拱门的图案需要多个基因在空间和时间上的精确表达，这通过分泌信号在组织之间进行协调。Wnt是在整个咽弓表达的分泌配体，但它们在颅面结构中的作用尚不清楚。在这项研究中，我们研究了Wnt在颅面图案中的作用，使用转基因斑马鱼抑制下游Wnt信号。我们发现，Wnt信号缺陷的胚胎具有下颌特异性缺陷，这与Bmp和Edn 1信号通路中的功能丧失表型非常相似。通过拯救实验，我们发现Wnt是Bmp和Edn 1信号的上游调节器。因此，我们已经发现了一个至关重要的要求Wnt信号在颅面图案。
Craniofacial development requires signals from epithelia to pattern skeletogenic neural crest (NC) cells, such as the subdivision of each pharyngeal arch into distinct dorsal (D) and ventral (V) elements. Wnt signaling has been implicated in many aspects of NC and craniofacial development, but its roles in D-V arch patterning remain unclear. To address this we blocked Wnt signaling in zebrafish embryos in a temporally-controlled manner, using transgenics to overexpress a dominant negative Tcf3, (dntcf3), (Tg(hsp70I:tcf3-GFP), or the canonical Wnt inhibitor dickkopf1 (dkk1), (Tg(hsp70i:dkk1-GFP) after NC migration. In dntcf3 transgenics, NC cells in the ventral arches of heat-shocked embryos show reduced proliferation, expression of ventral patterning genes (hand2, dlx3b, dlx5a, msxe), and ventral cartilage differentiation (e.g. lower jaws). These D-V patterning defects resemble the phenotypes of zebrafish embryos lacking Bmp or Edn1 signaling, and overexpression of dntcf3 dramatically reduces expression of a subset of Bmp receptors in the arches. Addition of ectopic BMP (or EDN1) protein partially rescues ventral development and expression of dlx3b, dlx5a, and msxe in Wnt signaling-deficient embryos, but surprisingly does not rescue hand2 expression. Thus Wnt signaling provides ventralizing patterning cues to arch NC cells, in part through regulation of Bmp and Edn1 signaling, but independently regulates hand2. Similarly, heat-shocked dkk1+ embryos exhibit ventral arch reductions, but also have mandibular clefts at the ventral midline not seen in dntcf3+ embryos. Dkk1 is expressed in pharyngeal endoderm, and cell transplantation experiments reveal that dntcf3 must be overexpressed in pharyngeal endoderm to disrupt D-V arch patterning, suggesting that distinct endodermal roles for Wnts and Wnt antagonists pattern the developing skeleton. Craniofacial abnormalities are among the most common birth defects. Understanding the molecular mechanisms underlying craniofacial disorders is crucial for developing treatment strategies. Much of the craniofacial skeleton arises from specialized embryonic structures known as pharyngeal arches. Patterning of these arches requires precise spatial and temporal expression of multiple genes, which is coordinated between tissues by secreted signals. Wnts are secreted ligands expressed throughout the pharyngeal arches yet their role in craniofacial patterning remains unclear. In this study we examine the role of Wnts in craniofacial patterning using transgenic zebrafish to inhibit downstream Wnt signaling. We show that Wnt signaling deficient embryos have lower jaw specific defects, which strongly resembles loss-of-function phenotypes in both the Bmp and Edn1 signaling pathways. Through rescue experiments we find that Wnts are upstream regulators of both Bmp and Edn1 signaling. We thus have uncovered a crucial requirement for Wnt signaling in craniofacial patterning.
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DOI：
10.1016/j.gep.2009.04.002
发表时间：
2009-06
期刊：
Gene expression patterns : GEP
影响因子：
--
作者：
Danesh SM;Villasenor A;Chong D;Soukup C;Cleaver O
通讯作者：
Cleaver O
影响因子：
2
作者：
Clouthier, David E.;Garcia, Elvin;Schilling, Thomas F.
通讯作者：
Schilling, Thomas F.
影响因子：
4.8
作者：
Filali, M;Cheng, NL;Engelhardt, JF
通讯作者：
Engelhardt, JF
DOI：
10.1073/pnas.0604751103
发表时间：
2006-08-01
影响因子：
11.1
作者：
Brito, Jose M.;Teillett, Marie-Aimee;Le Douarin, Nicole M.
通讯作者：
Le Douarin, Nicole M.
分析鞘氨醇1-磷酸信号传导突变体揭示了颌骨骨骼前体的生长，但没有背腹形图案。
影响因子：
2.7
作者：
Balczerski B;Matsutani M;Castillo P;Osborne N;Stainier DY;Crump JG
通讯作者：
Crump JG