Moody Sally A
Department of Anatomy and Regenerative Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA.
Methods Mol Biol. 2012;884:91-111. doi: 10.1007/978-1-61779-848-1_6.
The individual cells of the Xenopus cleavage-stage embryo have been fate mapped, revealing which of these cells contribute to the retina. Using this retina fate map, one can specifically modulate levels of gene expression in retina lineages to determine the function of proteins in various aspects of early retinal development, such as formation of the eye fields and determination of specific cell fates. This chapter presents the techniques for identifying specific retina blastomere precursor cells, and injecting them with lineage tracers, mRNAs encoding wild-type and mutant constructs or morpholino antisense oligonucleotides to alter gene expression.
非洲爪蟾卵裂期胚胎的单个细胞已进行了命运图谱绘制，揭示了这些细胞中哪些会发育成视网膜。利用这一视网膜命运图谱，可以特异性地调节视网膜谱系中的基因表达水平，以确定蛋白质在早期视网膜发育各个方面的功能，比如眼原基的形成以及特定细胞命运的决定。本章介绍了识别特定视网膜卵裂球前体细胞，并向其注射谱系示踪剂、编码野生型和突变体构建体的mRNA或吗啉代反义寡核苷酸以改变基因表达的技术。
