In vivo single-cell lineage tracing in zebrafish using high-resolution infrared laser-mediated gene induction microscopy

eLife

Xinwei Shen¹  Qiqi Sun¹  Shachuan Feng¹  Zilong Wen¹  Jianan Y Qu²  Kani Chen²  Xuesong Li²  Ye Tian³  Sicong He⁴  Yingzhu He⁵  Yi Wu⁵  Jin Xu⁵ 

[1] Center of Systems Biology and Human Health, The Hong Kong University of Science and Technology, Kowloon, China;Division of Life Science, The Hong Kong University of Science and Technology, Kowloon, China;State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Kowloon, China;Department of Electronic and Computer Engineering, The Hong Kong University of Science and Technology, Kowloon, China;State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Kowloon, China;

关键词: hematopoiesis;    lineage tracing;    single-cell labeling;    heat shock;   

DOI  :  10.7554/eLife.52024

来源: DOAJ

【 摘 要 】

Heterogeneity broadly exists in various cell types both during development and at homeostasis. Investigating heterogeneity is crucial for comprehensively understanding the complexity of ontogeny, dynamics, and function of specific cell types. Traditional bulk-labeling techniques are incompetent to dissect heterogeneity within cell population, while the new single-cell lineage tracing methodologies invented in the last decade can hardly achieve high-fidelity single-cell labeling and long-term in-vivo observation simultaneously. In this work, we developed a high-precision infrared laser-evoked gene operator heat-shock system, which uses laser-induced CreERT2 combined with loxP-DsRedx-loxP-GFP reporter to achieve precise single-cell labeling and tracing. In vivo study indicated that this system can precisely label single cell in brain, muscle and hematopoietic system in zebrafish embryo. Using this system, we traced the hematopoietic potential of hemogenic endothelium (HE) in the posterior blood island (PBI) of zebrafish embryo and found that HEs in the PBI are heterogeneous, which contains at least myeloid unipotent and myeloid-lymphoid bipotent subtypes.

【 授权许可】

Unknown