Zebrafish eaf1 and eaf2/u19 Mediate Effective Convergence and Extension Movements through the Maintenance of wnt11 and wnt5 Expression
Zebrafish eaf1 and eaf2/u19 Mediate Effective Convergence and Extension Movements through the Maintenance of wnt11 and wnt5 Expression
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斑马鱼 eaf1 和 eaf2/u19 通过维持 wnt11 和 wnt5 表达介导有效的收敛和伸展运动
DOI：
10.1074/jbc.m109.009654
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发表时间：
2009-06-12
影响因子：
4.8
通讯作者：
Xiao, Wuhan
中科院分区：
文献类型：
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作者：
Liu, Jing-Xia;Hu, Bo;Xiao, Wuhan
研究将 Eaf 家族的多种功能归因于 Eaf 家族，包括肿瘤抑制和眼睛发育。鉴于癌症与发育之间的潜在关联，我们开始利用斑马鱼探索 Eaf1 和 Eaf2/U19 在脊椎动物胚胎发生中的活性。原位杂交揭示了相似的 eaf1 和 eaf2/u19 表达模式。吗啡啉介导的 eaf1 或 eaf2/u19 表达敲低会产生类似的形态变化，这些变化可以通过靶标或相互靶标 mRNA 的异位表达来逆转。然而，Eaf1 和 Eaf2/U19 (Eafs)-吗啉代的组合增加了缺陷的严重性，表明 Eaf1 和 Eaf2/U19 仅共享一些功能冗余。 Eafs 敲低表型类似于在会聚和伸展运动方面存在缺陷的胚胎。事实上，敲低会导致收敛和延伸运动标记的表达模式发生变化，而使用 kaeda mRNA 的细胞追踪实验显示 Eafs 敲低和细胞迁移缺陷之间存在相关性。心脏和胰腺分化标志物显示，Eafs 敲低还破坏了心脏和胰腺器官前体的中线融合。非规范 Wnt 信号传导在器官前体的会聚和伸展运动以及中线会聚中发挥着关键作用。我们发现Eaf1和Eaf2/U19维持了wnt11和wnt5的表达水平。此外，wnt11或wnt5 mRNA部分挽救了eafs变形体中发生的会聚和伸展运动缺陷。 Wnt11 和 Wnt5 在 rhoA 上汇聚，因此毫不奇怪，rhoA mRNA 比单独使用 wnt11 或 wnt5 mRNA 更有效地修复缺陷。然而，wnt11和wnt5的异位表达并不影响eaf1和eaf2/u19的表达。这些数据表明 eaf1 和 eaf2/u19 在非规范 Wnt 信号传导的上游发挥作用，以介导收敛和延伸运动。
Studies have attributed several functions to the Eaf family, including tumor suppression and eye development. Given the potential association between cancer and development, we set forth to explore Eaf1 and Eaf2/U19 activity in vertebrate embryogenesis, using zebrafish. In situ hybridization revealed similar eaf1 and eaf2/u19 expression patterns. Morpholino-mediated knockdown of either eaf1 or eaf2/u19 expression produced similar morphological changes that could be reversed by ectopic expression of target or reciprocal-target mRNA. However, combination of Eaf1 and Eaf2/U19 (Eafs)-morpholinos increased the severity of defects, suggesting that Eaf1 and Eaf2/U19 only share some functional redundancy. The Eafs knockdown phenotype resembled that of embryos with defects in convergence and extension movements. Indeed, knockdown caused expression pattern changes for convergence and extension movement markers, whereas cell tracing experiments using kaeda mRNA showed a correlation between Eafs knockdown and cell migration defects. Cardiac and pancreatic differentiation markers revealed that Eafs knockdown also disrupted midline convergence of heart and pancreatic organ precursors. Noncanonical Wnt signaling plays a key role in both convergence and extension movements and midline convergence of organ precursors. We found that Eaf1 and Eaf2/U19 maintained expression levels of wnt11 and wnt5. Moreover, wnt11 or wnt5 mRNA partially rescued the convergence and extension movement defects occurring in eafs morphants. Wnt11 and Wnt5 converge on rhoA, so not surprisingly, rhoA mRNA more effectively rescued defects than either wnt11 or wnt5 mRNA alone. However, the ectopic expression of wnt11 and wnt5 did not affect eaf1 and eaf2/u19 expression. These data indicate that eaf1 and eaf2/u19 act upstream of noncanonical Wnt signaling to mediate convergence and extension movements.