In vivo analysis reveals a critical role for neuropilin-1 in cranial neural crest cell migration in chick
In vivo analysis reveals a critical role for neuropilin-1 in cranial neural crest cell migration in chick
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DOI：
10.1016/j.ydbio.2006.08.019
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发表时间：
2007-01-01
影响因子：
2.7
通讯作者：
Kulesa, Paul M.
中科院分区：
文献类型：
--
作者：
McLennan, Rebecca;Kulesa, Paul M.
神经嵴为研究侵袭性细胞迁移提供了一个优秀的模型系统，但目前尚不清楚分子机制如何以编程方式引导细胞到达精确的目标。我们研究了潜在的指导因子 Neuropilin-1 的作用，并使用功能性敲低测定、组织移植和体内共焦延时成像来分析鸡颅神经嵴细胞迁移模式的变化。当卵内的neuropilin-1功能被敲低时，神经嵴细胞无法完全侵入鳃弓，尤其是第二鳃弓。延时成像显示，neuropilin-1 siRNA 转染的神经嵴细胞在第二鳃弓入口处停止并塌陷丝状伪足，但并未死亡。这种表型是细胞自主的。为了测试群体压力和局部环境因素对驱动神经嵴细胞向鳃弓的影响，我们将 DiI 标记的神经嵴细胞的小亚群等时移植到去除了邻近的迁移前神经嵴细胞的宿主胚胎中。延时共聚焦分析表明，移植的细胞以狭窄、定向的流方式迁移。有趣的是，随着neuropilin-1功能的降低，神经嵴细胞仍然形成节段性迁移流，这表明最初的神经嵴细胞迁移和鳃弓的侵袭是分开的过程。 (c) 2006 Elsevier Inc. 保留所有权利。
The neural crest provides an excellent model system to study invasive cell migration, however it is still unclear how molecular mechanisms direct cells to precise targets in a programmed manner. We investigate the role of a potential guidance factor, neuropilin-1, and use functional knockdown assays, tissue transplantation and in vivo confocal time-lapse imaging to analyze changes in chick cranial neural crest cell migratory patterns. When neuropilin-1 function is knocked down in ovo, neural crest cells fail to fully invade the branchial arches, especially the 2nd branchial arch. Time-lapse imaging shows that neuropilin-1 siRNA transfected neural crest cells stop and collapse filopodia at the 2nd branchial arch entrances, but do not die. This phenotype is cell autonomous. To test the influence of population pressure and local environmental cues in driving neural crest cells to the branchial arches, we isochronically transplanted small subpopulations of DiI-labeled neural crest cells into host embryos ablated of neighboring, premigratory neural crest cells. Time-lapse confocal analysis reveals that the transplanted cells migrate in narrow, directed streams. Interestingly, with the reduction of neuropilin-1 function, neural crest cells still form segmental migratory streams, suggesting that initial neural crest cell migration and invasion of the branchial arches are separable processes. (c) 2006 Elsevier Inc. All rights reserved.