Shp2 knockdown and Noonan/LEOPARD mutant Shp2-induced gastrulation defects.
Shp2 knockdown and Noonan/LEOPARD mutant Shp2-induced gastrulation defects.
复制标题
SHP2敲低和NOONAN/LEOPARD突变体SHP2诱导的胃部缺陷。
DOI：
10.1371/journal.pgen.0030225
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发表时间：
2007-12
期刊：
影响因子：
4.5
通讯作者：
den Hertog, Jeroen
中科院分区：
文献类型：
--
作者：
Jopling, Chris;van Geemen, Daphne;den Hertog, Jeroen
Shp2是一种细胞质蛋白酪氨酸磷酸酶，对正常发育至关重要。在人类中已发现激活和失活突变分别引起相关的努南综合征和LEOPARD综合征。这些综合征的细胞生物学原因仍有待确定。我们使用斑马鱼来评估Shp2在早期发育中的作用。在这里，我们报告，吗啉介导的敲低斑马鱼Shp2导致原肠胚形成过程中的缺陷。细胞示踪实验表明，Shp2敲低诱导的收敛和伸展细胞运动的缺陷。使用一组标记物的原位杂交表明细胞命运不受Shp2敲低的影响。Shp2敲低诱导的缺陷被活性Fyn和Yes以及活性RhoA挽救。我们产生了Shp2的突变体，其突变在努南或LEOPARD综合征患者中鉴定，并确定努南Shp2被激活，LEOPARD Shp2缺乏催化蛋白酪氨酸磷酸酶活性。努南或LEOPARD突变体Shp2在斑马鱼胚胎中的表达诱导了会聚和伸展细胞运动缺陷，而不影响细胞命运。此外，这些胚胎表现出颅面和心脏缺陷，让人想起人类的症状。努南和LEOPARD突变体Shp2既不是加性的，也不是协同的，这与突变体Shp2在相同的信号传导途径中具有激活和失活作用一致。我们的研究结果表明，Shp2是所需的正常收敛和扩展细胞运动在原肠胚形成和Src家族激酶和RhoA的Shp2的下游。努南或LEOPARD Shp2的表达表型模拟了人类患者的颅面和心脏缺陷。Shp2信号缺陷早在原肠胚形成时就诱导细胞运动缺陷，这一发现可能对努南综合征和LEOPARD综合征的监测和诊断有意义。Shp2是一种蛋白质酪氨酸磷酸酶，Shp2的突变导致人类相关的努南综合征和LEOPARD综合征。我们使用斑马鱼研究Shp2在早期发育中的细胞生物学作用。Shp2敲低和突变Shp2的表达，其中含有对应于在人类努南和LEOPARD患者中发现的突变，诱导了原肠胚形成过程中相似的会聚和伸展细胞运动缺陷，而不影响细胞特化。活性Src家族激酶和活性RhoA挽救了Shp2敲低，表明Shp2下游的信号传导由Src家族激酶和RhoA介导。努南和LEOPARD Shp2在斑马鱼中的表达诱导了颅面和心脏缺陷，这让人联想到在人类患者中观察到的症状。共注射证明努南和LEOPARD Shp2不合作，这与两种突变体在相同的信号通路中起作用，具有相反的作用一致。Shp2信号缺陷早在原肠胚形成时就诱导细胞运动缺陷，这一发现可能对人类努南综合征和LEOPARD综合征的监测和诊断具有重要意义。
Shp2 is a cytoplasmic protein-tyrosine phosphatase that is essential for normal development. Activating and inactivating mutations have been identified in humans to cause the related Noonan and LEOPARD syndromes, respectively. The cell biological cause of these syndromes remains to be determined. We have used the zebrafish to assess the role of Shp2 in early development. Here, we report that morpholino-mediated knockdown of Shp2 in zebrafish resulted in defects during gastrulation. Cell tracing experiments demonstrated that Shp2 knockdown induced defects in convergence and extension cell movements. In situ hybridization using a panel of markers indicated that cell fate was not affected by Shp2 knock down. The Shp2 knockdown–induced defects were rescued by active Fyn and Yes and by active RhoA. We generated mutants of Shp2 with mutations that were identified in human patients with Noonan or LEOPARD Syndrome and established that Noonan Shp2 was activated and LEOPARD Shp2 lacked catalytic protein-tyrosine phosphatase activity. Expression of Noonan or LEOPARD mutant Shp2 in zebrafish embryos induced convergence and extension cell movement defects without affecting cell fate. Moreover, these embryos displayed craniofacial and cardiac defects, reminiscent of human symptoms. Noonan and LEOPARD mutant Shp2s were not additive nor synergistic, consistent with the mutant Shp2s having activating and inactivating roles in the same signaling pathway. Our results demonstrate that Shp2 is required for normal convergence and extension cell movements during gastrulation and that Src family kinases and RhoA were downstream of Shp2. Expression of Noonan or LEOPARD Shp2 phenocopied the craniofacial and cardiac defects of human patients. The finding that defective Shp2 signaling induced cell movement defects as early as gastrulation may have implications for the monitoring and diagnosis of Noonan and LEOPARD syndrome. Shp2 is a protein-tyrosine phosphatase and mutations in Shp2 cause the related Noonan and LEOPARD syndromes in humans. We used the zebrafish to investigate the cell biological role of Shp2 in early development. Shp2 knockdown and expression of mutant Shp2 that contained mutations corresponding to those found in human Noonan and LEOPARD patients, induced similar convergence and extension cell movement defects during gastrulation without affecting cell specification. Active Src family kinases and active RhoA rescued the Shp2 knockdown, indicating that signaling downstream of Shp2 was mediated by Src family kinases and RhoA. Expression of the Noonan and LEOPARD Shp2s in zebrafish induced craniofacial and cardiac defects that were reminiscent of the symptoms observed in human patients. Coinjections demonstrated that Noonan and LEOPARD Shp2s did not cooperate, which is consistent with the two mutants acting in the same signaling pathway with opposing effects. The finding that defective Shp2 signaling induced cell movement defects as early as gastrulation may have important implications for the monitoring and diagnosis of Noonan and LEOPARD syndromes in humans.
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