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
Further reading
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Adolescence is characterized by changes in reward-related behaviors, social behaviors, and decision-making. These behavioral changes are necessary for the transition into adulthood, but they also increase vulnerability to the development of a range of psychiatric disorders. Major reorganization of the dopamine system during adolescence is thought to underlie, in part, the associated behavioral changes and increased vulnerability. Here, we utilized fast scan cyclic voltammetry and microdialysis to examine differences in dopamine release as well as mechanisms that underlie differential dopamine signaling in the nucleus accumbens (NAc) core of adolescent (P28-35) and adult (P70-90) male rats. We show baseline differences between adult and adolescent-stimulated dopamine release in male rats, as well as opposite effects of the α6 nicotinic acetylcholine receptor (nAChR) on modulating dopamine release. The α6-selective blocker, α-conotoxin, increased dopamine release in early adolescent rats, but decreased dopamine release in rats beginning in middle adolescence and extending through adulthood. Strikingly, blockade of GABAA and GABAB receptors revealed that this α6-mediated increase in adolescent dopamine release requires NAc GABA signaling to occur. We confirm the role of α6 nAChRs and GABA in mediating this effect in vivo using microdialysis. Results herein suggest a multisynaptic mechanism potentially unique to the period of development that includes early adolescence, involving acetylcholine acting at α6-containing nAChRs to drive inhibitory GABA tone on dopamine release.
