Plessier Flora, Schmutz Sandrine, Novault Sophie, Marlow Heather
Committee on Development, Regeneration and Stem Cell Biology, The University of Chicago, Chicago, IL, USA.
École Doctorale "Complexité du Vivant" ED515, Sorbonne Université, Paris, France.
Methods Mol Biol. 2022;2450:565-581. doi: 10.1007/978-1-0716-2172-1_30.
Cnidarians have historically served as excellent laboratory models for regenerative development given their capacity to regrow large portions of the adult organism. This capacity is notably absent or poorly developed in the powerful genetic laboratory models Drosophila, C. elegans, and mouse. Increasingly, development of genetic and genomic resources and the application of next-generation sequencing-based techniques in cnidarian systems has further expanded the potential of cnidarian regenerative models. Here, we present a workflow for the characterization of the regenerative response in the sea anemone Nematostella vectensis utilizing fluorescence-activated cell sorting and a plate-based single-cell RNA-sequencing pipeline. This approach can characterize the transcriptional response during regeneration in distinct populations of cells, thus providing a quantitative view of a whole organism process at cellular resolution.
由于刺胞动物具有使成体生物的大部分组织再生的能力,它们长期以来一直是再生发育研究的优秀实验室模型。而在强大的遗传实验室模型果蝇、秀丽隐杆线虫和小鼠中,这种能力明显缺乏或发育不良。越来越多的遗传和基因组资源的开发以及基于下一代测序技术在刺胞动物系统中的应用,进一步扩展了刺胞动物再生模型的潜力。在此,我们展示了一种利用荧光激活细胞分选和基于平板的单细胞RNA测序流程来表征海葵星状海葵再生反应的工作流程。这种方法可以表征不同细胞群体在再生过程中的转录反应,从而在细胞分辨率上提供对整个生物体过程的定量观察。