In vivo single-cell lineage tracing in zebrafish using high-resolution infrared laser-mediated gene induction microscopy
- Hong Kong University of Science and Technology, Hong Kong
- South China University of Technology, China
Abstract
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.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figure 2-4, Figure 1-supplement 2, Figure 2-supplement 4, Figure 3-supplement 1, Figure 3-supplement 3, Figure 4-supplement 1.
Article and author information
Author details
Funding
Hong Kong University of Science and Technology (RPC10EG33)
- Jianan Y Qu
Research Grants Council, University Grants Committee (C6002-17GF)
- Jianan Y Qu
National Key R&D Program of China (2018YFA0800200)
- Jin Xu
Research Grants Council, University Grants Committee (662513)
- Jianan Y Qu
Research Grants Council, University Grants Committee (16103215)
- Jianan Y Qu
Research Grants Council, University Grants Committee (16148816)
- Jianan Y Qu
Research Grants Council, University Grants Committee (16102518)
- Jianan Y Qu
Research Grants Council, University Grants Committee (T13-607/12R)
- Jianan Y Qu
Research Grants Council, University Grants Committee (T13-706/11-1)
- Jianan Y Qu
Research Grants Council, University Grants Committee (AOE/M-09/12)
- Jianan Y Qu
Research Grants Council, University Grants Committee (T13-605/18W)
- Jianan Y Qu
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2020, He et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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In vivo single-cell lineage tracing in zebrafish using high-resolution infrared laser-mediated gene induction microscopy
eLife 9:e52024.
https://doi.org/10.7554/eLife.52024
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