Abstract
The lymphatic system has become the subject of great interest in recent years because of its important role in normal and pathological processes. Progress in understanding the origins and early development of this system, however, has been hampered by difficulties in observing lymphatic cells in vivo and in performing defined genetic and experimental manipulation of the lymphatic system in currently available model organisms. Here, we show that the optically clear developing zebrafish provides a useful model for imaging and studying lymphatic development, with a lymphatic system that shares many of the morphological, molecular and functional characteristics of the lymphatic vessels found in other vertebrates. Using two-photon time-lapse imaging of transgenic zebrafish, we trace the migration and lineage of individual cells incorporating into the lymphatic endothelium. Our results show lymphatic endothelial cells of the thoracic duct arise from primitive veins through a novel and unexpected pathway.
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Acknowledgements
The authors would like to thank W.-K. Chan (National University of Singapore) for providing the prox1 plasmid; M. Klagsbrun (Harvard University) for the nrp2a plasmid, B. Roman (Georgetown University) for the Tg(fli1:EGFP)yl, Tg(gata1:dsRed) fish, and K. Noma, Y. Takahashi and the members of the bio-imaging center of Iwate Medical University for technical assistance. We are grateful to all the members of the Weinstein lab for discussions and technical assistance. We also thank G. Oliver for discussions and I.B. Dawid for critical reading of this manuscript. K.Y. is supported by an EMBO long-term fellowship. B.M.W. is supported by the intramural program of the US National Institutes of Health's National Institute of Child Health and Human Development.
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Supplementary information
Supplementary Fig. 1
Assembly of the developing thoracic duct. (PDF 194 kb)
Supplementary Fig. 2
The lymphatic thoracic duct is present in the adult zebrafish trunk. (PDF 279 kb)
Supplementary Fig. 3
Molecular characterization of the thoracic duct. (PDF 212 kb)
Supplementary Fig. 4
Emergence of buds for the jugular lymphatics from the common cardinal vein, at or near the future drainage point. (PDF 179 kb)
Supplementary Video 1
This movie shows two-photon time-lapse images of a thoracic duct sprout in the ventral trank of a Tg(fli:EGFP)y1 transgenic zebrafish. (MOV 10148 kb)
Supplementary Video 2
This movie shows two-photon time-lapse images of endothelial cell nuclear dynamics in Tg(fli:EGFP)y1 transgenic zebrafish. (MOV 9235 kb)
Supplementary Video 3
This movie shows two-photon time-lapse images of endothelial cell nuclear dynamics in Tg(fli:EGFP)y1 transgenic zebrafish. (MOV 8624 kb)
Supplementary Video 4
This is a previously published movie from Isogai, et al. (Development 130, 5281; 2003) reproduced here for the convenience of the reader. (MOV 984 kb)
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Yaniv, K., Isogai, S., Castranova, D. et al. Live imaging of lymphatic development in the zebrafish. Nat Med 12, 711–716 (2006). https://doi.org/10.1038/nm1427
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DOI: https://doi.org/10.1038/nm1427
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