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.
Reviewing Editor
- Philippe Herbomel, Institut Pasteur, CNRS UMR3738, France
Version history
- Received: September 20, 2019
- Accepted: January 4, 2020
- Accepted Manuscript published: January 6, 2020 (version 1)
- Version of Record published: February 13, 2020 (version 2)
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
Further reading
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Large-scale cell flow characterizes gastrulation in animal development. In amniote gastrulation, particularly in avian gastrula, a bilateral vortex-like counter-rotating cell flow, called ‘polonaise movements’, appears along the midline. Here, through experimental manipulations, we addressed relationships between the polonaise movements and morphogenesis of the primitive streak, the earliest midline structure in amniotes. Suppression of the Wnt/planar cell polarity (PCP) signaling pathway maintains the polonaise movements along a deformed primitive streak. Mitotic arrest leads to diminished extension and development of the primitive streak and maintains the early phase of the polonaise movements. Ectopically induced Vg1, an axis-inducing morphogen, generates the polonaise movements, aligned to the induced midline, but disturbs the stereotypical cell flow pattern at the authentic midline. Despite the altered cell flow, induction and extension of the primitive streak are preserved along both authentic and induced midlines. Finally, we show that ectopic axis-inducing morphogen, Vg1, is capable of initiating the polonaise movements without concomitant PS extension under mitotic arrest conditions. These results are consistent with a model wherein primitive streak morphogenesis is required for the maintenance of the polonaise movements, but the polonaise movements are not necessarily responsible for primitive streak morphogenesis. Our data describe a previously undefined relationship between the large-scale cell flow and midline morphogenesis in gastrulation.
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- Developmental Biology
- Physics of Living Systems
Geometric criteria can be used to assess whether cell intercalation is active or passive during the convergent extension of tissue.
