Abstract:
Development and regeneration occur by a process of genetically encoded spatiotemporally dynamic cellular interactions. The use of cell transplantation between animals to track cell fate and to induce mismatches in the genetic, spatial, or temporal properties of donor and host cells is a powerful means of examining the nature of these interactions. Organisms such as chick and amphibians have made crucial contributions to our understanding of development and regeneration, respectively, in large part because of their amenability to transplantation. The power of these models, however, has been limited by low genetic tractability. Likewise, the major genetic model organisms have lower amenability to transplantation. The zebrafish is a major genetic model for development and regeneration, and while cell transplantation is common in zebrafish, it is generally limited to the transfer of undifferentiated cells at the early blastula and gastrula stages of development. In this article, we present a simple and robust method that extends the zebrafish transplantation window to any embryonic or larval stage between at least 1 and 7 days post fertilization. The precision of this approach allows for the transplantation of as little as one cell with near-perfect spatial and temporal resolution in both donor and host animals. While we highlight here the transplantation of embryonic and larval neurons for the study of nerve development and regeneration, respectively, this approach is applicable to a wide range of progenitor and differentiated cell types and research questions.
摘要:
发育和再生通过基因编码的时空动态细胞相互作用的过程发生。利用动物之间的细胞移植来追踪细胞命运并诱导遗传上的错配,空间,或供体和宿主细胞的时间特性是检查这些相互作用性质的有力手段。小鸡和两栖动物等生物对我们对发育和再生的理解做出了至关重要的贡献,分别,很大程度上是因为它们对移植的适应性。这些模型的力量,然而,受到低遗传可操作性的限制。同样,主要的遗传模式生物对移植的适应性较低。斑马鱼是发育和再生的主要遗传模子,虽然细胞移植在斑马鱼中很常见,它通常仅限于在早期囊胚和原肠胚发育阶段的未分化细胞的转移。在这篇文章中,我们提出了一种简单而强大的方法,可以将斑马鱼移植窗口扩展到受精后至少1到7天之间的任何胚胎或幼虫阶段。这种方法的精度允许在供体和宿主动物中移植少至一个具有近乎完美的空间和时间分辨率的细胞。虽然我们在这里强调胚胎和幼虫神经元的移植,以研究神经发育和再生,分别,这种方法适用于广泛的祖细胞和分化细胞类型和研究问题。