title: Multicolor Cell Barcoding Technology for Long-Term Surveillance of Epithelial Regeneration in Zebrafish.
abstract: 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.
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Chen-Hui
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Laboratory of Cell Migration, Candiolo Cancer Institute - FPO, IRCCS, Candiolo, Torino 10060, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Puliafito
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Ben D
authors.initials: BD
authors.lastname: Cox
authors.affiliation: Laboratory of Cell Migration, Candiolo Cancer Institute - FPO, IRCCS, Candiolo, Torino 10060, Italy; Department of Oncology, University of Torino, Torino 10060, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Primo
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Stefano
authors.initials: S
authors.lastname: Di Talia
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA. Electronic address: kenneth.poss@duke.edu.
authors.forename: Kenneth D
authors.initials: KD
authors.lastname: Poss
mesh_terms: D058500:Animal Fins / Q000293:injuries / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D004847:Epithelial Cells / Q000502:physiology; D004848:Epithelium / Q000293:injuries / Q000502:physiology; D006706:Homeostasis; D023421:Models, Animal; D017382:Reactive Oxygen Species / Q000378:metabolism; D012038:Regeneration / Q000235:genetics / Q000502:physiology; D012867:Skin / Q000293:injuries; D012880:Skin Pigmentation / Q000235:genetics / Q000502:physiology; D015027:Zebrafish / Q000235:genetics* / Q000502:physiology*
chemical_list: D017382:Reactive Oxygen Species