Phylogenetic fate mapping
Phylogenetic fate mapping
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DOI：
10.1073/pnas.0601265103
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发表时间：
2006-04-04
影响因子：
11.1
通讯作者：
Horwitz, MS
中科院分区：
文献类型：
--
作者：
Salipante, SJ;Horwitz, MS
细胞命运图描述了细胞分裂、迁移和凋亡的顺序如何将受精卵转化为成体。然而，只有在秀丽隐杆线虫中，对每个细胞分裂的显微镜观察才允许构建一个完整的命运图。更复杂、更不透明的动物被证明不那么屈服。然而，DNA复制会产生体细胞突变。因此，多细胞生物由马赛克组成，其中大多数细胞获得独特的基因组，这些基因组可能能够描绘它们的祖先。在这里，我们采取一种系统发育的方法，通过推断在发育过程中突变发生的顺序，被动地追溯胚胎关系。我们发现多鸟嘌呤重复DNA序列是特别有用的遗传标记，因为它们在有丝分裂期间经常改变长度。为了证明可行性，我们基于影响多鸟嘌呤标记长度的突变，从系统发育上重建了培养小鼠NIH 3T3细胞的谱系。然后，我们采用全基因组扩增对取自小鼠的单细胞中的多鸟嘌呤标记物进行基因型分析，并使用系统遗传学来推断样本组织的发育关系。结果与目前对胚胎发生的理解一致，并证明了这种方法在单细胞分辨率下产生完整哺乳动物细胞命运的大规模潜力。
Cell fate maps describe how the sequence of cell division, migration, and apoptosis transform a zygote into an adult. Yet, it is only in Caenorhabditis elegans where microscopic observation of each cell division has allowed for construction of a complete fate map. More complex, and opaque, animals prove less yielding. DNA replication, however, generates somatic mutations. Consequently, multicellular organisms comprise mosaics where most cells acquire unique genomes that are potentially capable of delineating their ancestry. Here we take a phylogenetic approach to passively retrace embryonic relationships by deducing the order in which mutations have arisen during development. We show that polyguanine repeat DNA sequences are particularly useful genetic markers, because they frequently change length during mitosis. To demonstrate feasibility, we phylogenetically reconstruct the lineage of cultured mouse NIH 3T3 cells based on mutations affecting the length of polyguanine markers. We then employ whole genome amplification to genotype polyguanine markers in single cells taken from a mouse and use phylogenetics to infer the developmental relationships of the sampled tissues. The result is consistent with the present understanding of embryogenesis and demonstrates the large scale potential of this method for producing a complete mammalian cell fate at the resolution of a single cell.