Gallardo Viviana E, Liang Jin, Behra Martine, Elkahloun Abdel, Villablanca Eduardo J, Russo Vincenzo, Allende Miguel L, Burgess Shawn M
Center for Genome Regulation. Facultad de Ciencias, Universidad de Chile, Casilla 653. Santiago, Chile.
BMC Dev Biol. 2010 Dec 13;10:120. doi: 10.1186/1471-213X-10-120.
Development of the posterior lateral line (PLL) system in zebrafish involves cell migration, proliferation and differentiation of mechanosensory cells. The PLL forms when cranial placodal cells delaminate and become a coherent, migratory primordium that traverses the length of the fish to form this sensory system. As it migrates, the primordium deposits groups of cells called neuromasts, the specialized organs that contain the mechanosensory hair cells. Therefore the primordium provides both a model for studying collective directional cell migration and the differentiation of sensory cells from multipotent progenitor cells.
Through the combined use of transgenic fish, Fluorescence Activated Cell Sorting and microarray analysis we identified a repertoire of key genes expressed in the migrating primordium and in differentiated neuromasts. We validated the specific expression in the primordium of a subset of the identified sequences by quantitative RT-PCR, and by in situ hybridization. We also show that interfering with the function of two genes, f11r and cd9b, defects in primordium migration are induced. Finally, pathway construction revealed functional relationships among the genes enriched in the migrating cell population.
Our results demonstrate that this is a robust approach to globally analyze tissue-specific expression and we predict that many of the genes identified in this study will show critical functions in developmental events involving collective cell migration and possibly in pathological situations such as tumor metastasis.
斑马鱼后侧线(PLL)系统的发育涉及机械感觉细胞的细胞迁移、增殖和分化。当颅基板细胞分层并形成一个连贯的、迁移的原基时,PLL形成,该原基贯穿鱼的全长以形成这个感觉系统。在迁移过程中,原基会沉积称为神经丘的细胞群,神经丘是包含机械感觉毛细胞的特殊器官。因此,原基既为研究集体定向细胞迁移提供了模型,也为研究多能祖细胞分化为感觉细胞提供了模型。
通过联合使用转基因鱼、荧光激活细胞分选和微阵列分析,我们确定了在迁移的原基和分化的神经丘中表达的关键基因库。我们通过定量RT-PCR和原位杂交验证了所鉴定序列的一个子集在原基中的特异性表达。我们还表明,干扰两个基因f11r和cd9b的功能会诱导原基迁移缺陷。最后,通路构建揭示了迁移细胞群体中富集的基因之间的功能关系。
我们的结果表明,这是一种全面分析组织特异性表达的有效方法,并且我们预测本研究中鉴定的许多基因将在涉及集体细胞迁移的发育事件中以及可能在诸如肿瘤转移等病理情况下发挥关键作用。