Wnt/β-catenin signaling was disrupted by deleting supraorbital mesenchymal β-catenin or epithelial Wls. p63 was removed to determine whether the expression of Wnts is affected. The eyelid morphology was examined at different stages. Proliferation, apoptosis, and expression of Wnt ligands and their target genes were analyzed via immunofluorescence staining, TUNEL assay, and in situ hybridization.
Deletion of β-catenin in supraorbital mesenchyme abolishes eyelid growth by causing decreased proliferation in supraorbital epithelium and underlying mesenchyme. Inhibition of Wnt secretion by deleting Wls in supraorbital epithelium results in failure of eyelid development, similar to the effects of deleting mesenchymal β-catenin. Knockout of p63 results in formation of hypoplastic eyelids and reduced expression of several Wnt ligands in eyelid epithelium.
Epithelial Wnt ligands activate mesenchymal Wnt/β-catenin signaling to control eyelid growth and their expression is partially regulated by p63.
方法:通过删除眶上间充质β-catenin或上皮Wls来破坏Wnt/β-catenin信号传导。去除p63以确定Wnts的表达是否受到影响。在不同阶段检查眼睑形态。扩散,凋亡,并通过免疫荧光染色分析Wnt配体及其靶基因的表达,TUNEL检测,和原位杂交。
结果:眶上间充质中β-连环蛋白的缺失通过减少眶上上皮和下层间充质的增殖来消除眼睑生长。通过删除眶上上皮中的Wls来抑制Wnt分泌导致眼睑发育失败,与删除间充质β-catenin的效果相似。p63基因敲除导致眼睑发育不良的形成和眼睑上皮中几种Wnt配体的表达减少。
结论:上皮Wnt配体激活间充质Wnt/β-catenin信号传导以控制眼睑生长,其表达部分受p63调控。