Hu Diane, Marcucio Ralph S
Department of Orthopaedic Surgery, University of California, San Francisco, CA, USA.
J Vis Exp. 2011 Mar 24(49):2557. doi: 10.3791/2557.
The accessibility of avian embryos has helped experimental embryologists understand the fates of cells during development and the role of tissue interactions that regulate patterning and morphogenesis of vertebrates (e.g., (1, 2, 3, 4)). Here, we illustrate a method that exploits this accessibility to test the signaling and patterning properties of ectodermal tissues during facial development. In these experiments, we create quail-chick (5) or mouse-chick (6) chimeras by transplanting the surface cephalic ectoderm that covers the upper jaw from quail or mouse onto either the same region or an ectopic region of chick embryos. The use of quail as donor tissue for transplantation into chicks was developed to take advantage of a nucleolar marker present in quail but not chick cells, thus allowing investigators to distinguish host and donor tissues (7). Similarly, a repetitive element is present in the mouse genome and is expressed ubiquitously, which allows us to distinguish host and donor tissues in mouse-chick chimeras (8). The use of mouse ectoderm as donor tissue will greatly extend our understanding of these tissue interactions, because this will allow us to test the signaling properties of ectoderm derived from various mutant embryos.
鸟类胚胎易于获取，这有助于实验胚胎学家了解发育过程中细胞的命运以及调节脊椎动物模式形成和形态发生的组织相互作用的作用（例如，(1, 2, 3, 4)）。在这里，我们阐述一种利用这种易获取性来测试面部发育过程中外胚层组织的信号传导和模式形成特性的方法。在这些实验中，我们通过将覆盖鹌鹑或小鼠上颌的头部表面外胚层移植到鸡胚胎的相同区域或异位区域来创建鹌鹑 - 鸡（5）或小鼠 - 鸡（6）嵌合体。将鹌鹑用作移植到鸡中的供体组织是为了利用鹌鹑细胞中存在而鸡细胞中不存在的核仁标记，从而使研究人员能够区分宿主和供体组织（7）。同样，小鼠基因组中存在一种重复元件且普遍表达，这使我们能够在小鼠 - 鸡嵌合体中区分宿主和供体组织（8）。使用小鼠外胚层作为供体组织将极大地扩展我们对这些组织相互作用的理解，因为这将使我们能够测试源自各种突变胚胎的外胚层的信号传导特性。
