Carroll Shannon H, Schafer Sogand, Kawasaki Kenta, Tsimbal Casey, Julé Amélie M, Hallett Shawn A, Li Edward, Liao Eric C
Center for Craniofacial Innovation, Children's Hospital of Philadelphia Research Institute, Children's Hospital of Philadelphia, PA 19104, USA.
Division of Plastic and Reconstructive Surgery, Department of Surgery, Children's Hospital of Philadelphia, PA 19104, USA.
bioRxiv. 2024 Sep 4:2023.11.07.566024. doi: 10.1101/2023.11.07.566024.
Wnt signaling plays crucial roles in embryonic patterning including the regulation of convergent extension during gastrulation, the establishment of the dorsal axis, and later, craniofacial morphogenesis. Further, Wnt signaling is a crucial regulator of craniofacial morphogenesis. The adapter proteins Dact1 and Dact2 modulate the Wnt signaling pathway through binding to Disheveled. However, the distinct relative functions of Dact1 and Dact2 during embryogenesis remain unclear. We found that and genes have dynamic spatiotemporal expression domains that are reciprocal to one another suggesting distinct functions during zebrafish embryogenesis. Both and contribute to axis extension, with compound mutants exhibiting a similar convergent extension defect and craniofacial phenotype to the mutant. Utilizing single-cell RNAseq and an established noncanonical Wnt pathway mutant with a shortened axis (), we identified specific roles during early development. Comparative whole transcriptome analysis between wildtype and and wildtype and compound mutants revealed a novel role for in regulating the mRNA expression of the classical calpain . Over-expression of phenocopies craniofacial dysmorphology. These results identify a previously unappreciated role of and calcium-dependent proteolysis during embryogenesis. Taken together, our findings highlight the distinct and overlapping roles of and in embryonic craniofacial development, providing new insights into the multifaceted regulation of Wnt signaling.
Wnt信号通路在胚胎模式形成中发挥着关键作用，包括在原肠胚形成过程中对汇聚延伸的调控、背轴的建立，以及随后的颅面形态发生。此外，Wnt信号通路是颅面形态发生的关键调节因子。衔接蛋白Dact1和Dact2通过与Disheveled结合来调节Wnt信号通路。然而，Dact1和Dact2在胚胎发生过程中各自独特的相对功能仍不清楚。我们发现，[基因名称1]和[基因名称2]基因具有动态的时空表达域，它们彼此相反，这表明在斑马鱼胚胎发生过程中具有不同的功能。[基因名称1]和[基因名称2]都对轴延伸有贡献，复合突变体表现出与[基因名称1]突变体相似的汇聚延伸缺陷和颅面表型。利用单细胞RNA测序和一个已建立的轴缩短的非经典Wnt通路突变体（[突变体名称]），我们确定了[基因名称1]在早期发育中的特定作用。野生型与[基因名称1]以及野生型与[基因名称1][基因名称2]复合突变体之间的比较全转录组分析揭示了[基因名称1]在调节经典钙蛋白酶[蛋白酶名称]的mRNA表达方面的新作用。[基因名称1]的过表达模拟了[基因名称1]的颅面畸形。这些结果确定了[基因名称1]和钙依赖性蛋白水解在胚胎发生过程中以前未被认识到的作用。综上所述，我们的发现突出了[基因名称1]和[基因名称2]在胚胎颅面发育中的不同和重叠作用，为Wnt信号通路的多方面调节提供了新的见解。
