Multicolor Cell Barcoding Technology for Long-Term Surveillance of Epithelial Regeneration in Zebrafish

• Published in: Developmental cell Vol. 36; no. 6; pp. 668 - 680
• Main Authors: Chen, Chen-Hui, Puliafito, Alberto, Cox, Ben D., Primo, Luca, Fang, Yi, Di Talia, Stefano, Poss, Kenneth D.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21.03.2016
• Subjects: Animal Fins - injuries; Animal Fins - physiology; Animals; Animals, Genetically Modified; DNA Barcoding, Taxonomic - methods; Epithelial Cells - physiology; Epithelium - injuries; Epithelium - physiology; Homeostasis; Models, Animal; Reactive Oxygen Species - metabolism; Regeneration - genetics; Regeneration - physiology; Skin - injuries; Skin Pigmentation - genetics; Skin Pigmentation - physiology; Zebrafish - genetics; Zebrafish - physiology
• ISSN: 1534-5807; 1878-1551
• DOI: 10.1016/j.devcel.2016.02.017

Current fate mapping and imaging platforms are limited in their ability to capture dynamic behaviors of epithelial cells. To deconstruct regenerating adult epithelial tissue at single-cell resolution, we created a multicolor system, skinbow, that barcodes the superficial epithelial cell (SEC) population of zebrafish skin with dozens of distinguishable tags. With image analysis to directly segment and simultaneously track hundreds of SECs in vivo over entire surface lifetimes, we readily quantified the orchestration of cell emergence, growth, repositioning, and loss under homeostatic conditions and after exfoliation or appendage amputation. We employed skinbow-based imaging in conjunction with a live reporter of epithelial stem cell cycle activity and as an instrument to evaluate the effects of reactive oxygen species on SEC behavior during epithelial regeneration. Our findings introduce a platform for large-scale, quantitative in vivo imaging of regenerating skin and reveal unanticipated collective dynamism in epithelial cell size, mobility, and interactions. [Display omitted] •Transgenic skinbow zebrafish barcodes skin superficial epithelial cells (SECs)•Hundreds of SECs are simultaneously monitored in vivo over cell surface lifetimes•SEC behaviors quantified during regeneration of skin wounds or amputation injuries•Identification and quantification of ROS effects on SEC regeneration Chen et al. present a multicolor transgenic system in adult zebrafish for simultaneous tracking of hundreds of individual surface epithelial cells over long time periods. They identify and quantify collective cell behaviors enabling skin homeostatic maintenance, and they directly visualize and quantify at single-cell resolution injury impacts on epithelial cells.