通过卵裂球缺失、移植和外植体培养来检测非洲爪蟾的视网膜命运决定。

Testing retina fate commitment in Xenopus by blastomere deletion, transplantation, and explant culture.

作者信息

Moody Sally A

机构信息

Department of Anatomy and Regenerative Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA.

出版信息

Methods Mol Biol. 2012;884:115-27. doi: 10.1007/978-1-61779-848-1_7.

DOI:10.1007/978-1-61779-848-1_7

PMID:22688701

Abstract

The lineages of individual cells of the Xenopus cleavage-stage embryo have been fate-mapped to reveal the subset of blastomeres that are the major and minor precursors of the retina. Using this retina fate map, one can test the commitment of each of these cells to various retinal cell fates by manipulating the environment in which they develop. This chapter presents the techniques for identifying specific retina blastomere precursor cells, deleting them to test whether they are required for producing specific kinds of retinal cells, transplanting them to novel embryonic locations in host embryos to test whether they are committed to produce specific kinds of retinal cells, and growing them in explant culture to determine if their ability to produce specific kinds of retinal cells is autonomous.

摘要

非洲爪蟾卵裂期胚胎单个细胞的谱系已通过命运图谱绘制出来，以揭示作为视网膜主要和次要前体的卵裂球子集。利用这个视网膜命运图谱，可以通过操纵细胞发育的环境来测试这些细胞中每一个对各种视网膜细胞命运的定向。本章介绍了识别特定视网膜卵裂球前体细胞的技术，删除这些细胞以测试产生特定类型视网膜细胞是否需要它们，将它们移植到宿主胚胎的新胚胎位置以测试它们是否注定会产生特定类型的视网膜细胞，以及在体外培养中培养它们以确定它们产生特定类型视网膜细胞的能力是否是自主的。