Endo Yukinori
Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Curr Protoc Cell Biol. 2012 Sep;Chapter 19:Unit19.15. doi: 10.1002/0471143030.cb1915s56.
Important events in embryonic development such as gastrulation, neurulation, and cranial neural crest development occur in ectodermal tissues during vertebrate embryonic development. Although the chicken embryo is a well-established model system in developmental biology, problems of accessibility of the ectoderm for experimental manipulation and an inability to generate gene knockouts previously impeded studies of gene regulation and key processes during chicken gastrulation and neurulation. The technique of in ovo electroporation permits genetic manipulation and provides a powerful animal model. However, the problem of accessibility to the ectoderm in ovo requires an ex ovo whole-embryo culture approach combined with electroporation. This unit provides convenient and reproducible whole-embryo ex ovo culture and electroporation protocols. These chicken embryo culture protocols can be used not only for gene regulatory experiments, but also for time-lapse imaging of the dynamics of early vertebrate development.
在脊椎动物胚胎发育过程中，诸如原肠胚形成、神经胚形成和颅神经嵴发育等胚胎发育中的重要事件发生在外胚层组织中。尽管鸡胚是发育生物学中一个成熟的模型系统，但外胚层难以进行实验操作以及先前无法产生基因敲除的问题，阻碍了对鸡原肠胚形成和神经胚形成过程中的基因调控及关键过程的研究。卵内电穿孔技术允许进行基因操作，并提供了一个强大的动物模型。然而，卵内获取外胚层的问题需要结合电穿孔的卵外全胚胎培养方法。本单元提供了方便且可重复的全胚胎卵外培养和电穿孔方案。这些鸡胚培养方案不仅可用于基因调控实验，还可用于早期脊椎动物发育动态的延时成像。
