Max-Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
Dev Biol. 2013 Jan 1;373(1):196-204. doi: 10.1016/j.ydbio.2012.10.019. Epub 2012 Oct 24.
Limb regeneration involves re-establishing a limb development program from cells within adult tissues. Identifying molecular handles that provide insight into the relationship between cell differentiation status and cell lineage is an important step to study limb blastema cell formation. Here, using single cell PCR, focusing on newly isolated Twist1 sequences, we molecularly profile axolotl limb blastema cells using several progenitor cell markers. We link their molecular expression profile to their embryonic lineage via cell tracking experiments. We use in situ hybridization to determine the spatial localization and extent of overlap of different markers and cell types. Finally, we show by single cell PCR that the mature axolotl limb harbors a small but significant population of Twist1(+) cells.
肢体再生涉及到从成年组织中的细胞重新建立肢体发育程序。确定分子处理方法,深入了解细胞分化状态和细胞谱系之间的关系,是研究肢体芽基细胞形成的重要步骤。在这里,我们使用单细胞 PCR,重点关注新分离的 Twist1 序列,使用几种祖细胞标记物对蝾螈肢体芽基细胞进行分子特征分析。我们通过细胞追踪实验将它们的分子表达谱与其胚胎谱系联系起来。我们使用原位杂交技术来确定不同标记物和细胞类型的空间定位和重叠程度。最后,我们通过单细胞 PCR 表明,成熟的蝾螈肢体中存在一小部分但具有重要意义的 Twist1(+)细胞。