Journal
DEVELOPMENTAL DYNAMICS
Volume 250, Issue 5, Pages 701-716Publisher
WILEY
DOI: 10.1002/dvdy.286
Keywords
3D imaging; infectiology; lymphatic network; tissue clearing; zebrafish phenotyping
Categories
Funding
- Agence Nationale de la Recherche [Phylogen DC ANR-09-BLAN-0073-02]
- Projet TEFOR-Investissement d'avenir ANR-II-INBS
- FP7 People: Marie-Curie Actions [FishForPharma PITN-GA-2011-289209]
- H2020 European Institute of Innovation and Technology [Vetbionet 731014]
- INRAE
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In zebrafish, lymphatic endothelial cells (LECs) originate from multiple distinct progenitor populations and form organ-specific lymphatic vasculatures. A new transgenic line expressing eGFP under the control of zebrafish batf3 promoter has been established, providing a useful system to study LEC populations with heterogeneity depending on progenitor origin, tissue environment, and physiological conditions. Additionally, a novel fish-adapted tissue clearing method has been developed for deep imaging and 3D-visualization of vascular and lymphatic networks in the whole organism.
Background: In zebrafish, lymphatic endothelial cells (LECs) originate from multiple/several distinct progenitor populations and generate organ-specific lymphatic vasculatures. Cell fate and tissue specificities were determined using a combination of genetically engineered transgenic lines in which the promoter of a LEC-specific gene drives expression of a fluorescent reporter protein. Results: We established a novel zebrafish transgenic line expressing eGFP under the control of part of the zebrafish batf3 promoter (Basic Leucine Zipper ATF-Like Transcription Factor 3). Spatiotemporal examination of Tg(batf3MIN:eGFP) transgenic fish revealed a typical lymphatic expression pattern, which does not perfectly recapitulate the expression pattern of existing LEC transgenic lines. eGFP(+) cells constitute a heterogeneous endothelial cell population, which expressed LEC and/or blood endothelial cells (BEC) markers in different tissues. In addition, we characterize the renal eGFP(+) cell as a population of interest to study kidney diseases and regeneration. Conclusion: Our Tg(batf3MIN:eGFP) reporter zebrafish line provides a useful system to study LEC populations, of which heterogeneity depends on origin of progenitors, tissue environment and physiological conditions. We further developed a novel fish-adapted tissue clearing method, which allows deep imaging and 3D-visualization of vascular and lymphatic networks in the whole organism.
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