A transgenic Xenopus laevis reporter product to study lymphangiogenesis [Analysis Report]
- Annelii Ny1,2,*,
- Wouter Vandevelde1,2,*,
- Philipp Hohensinnerone,2,
- Manu Beerens3,
- Ilse Geudensone,2,
- Antonio Diez-Juan1,2,‡,
- Katleen Brepoels1,2,
- Stéphane Plaisanceone,2,
- Paul A. Krieg4,
- Tobias Langenbergone,two,
- Stefan Vinckierone,2,
- Aernout Luttun3,
- Peter Carmelietone,2 and
- Mieke Dewerchinone,two,§
oneLaboratory of Angiogenesis and Neurovascular hyperlink, Vesalius Study Center, VIB, 3000 Leuven, Belgium
twoLaboratory of Angiogenesis and Neurovascular url, Vesalius Research Center, KU Leuven, 3000 Leuven, Belgium
threeCentre for Molecular and Vascular Biology, KU Leuven, 3000 Leuven, Belgium
fourDepartment of Mobile Biology and Anatomy, College of Arizona School of Medicine, Tucson, AZ 85724-5044, Usa
‡Current deal with: Instituto de Investigación Sanitaria INCLIVA, 46010 Valencia, Spain
- ↵§Author for correspondence ( )
↵* These authors contributed equally to this operate
The value of the blood- and lymph vessels in the transport of crucial fluids, gases, macromolecules and cells in vertebrates
warrants best insight into the regulatory mechanisms underlying their advancement. Mouse and zebrafish designs of lymphatic
development are instrumental for gene discovery and gene characterization but are tough for specified factors, e.g. no
immediate accessibility of embryonic levels, or non-straightforward visualization of early lymphatic sprouting, respectively.
We previously shown that the Xenopus tadpole is a worthwhile model to review the procedures of lymphatic advancement. Nonetheless, a fluorescent Xenopus reporter immediately visualizing the lymph vessels was lacking. Here, we produced transgenic Tg(Flk1:eGFP) Xenopus laevis reporter strains expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter.
We also recognized a large-resolution fluorescent dye labeling approach selectively and persistently visualizing lymphatic
endothelial cells, even in circumstances of impaired lymph vessel formation or drainage perform upon silencing of lymphangiogenic
factors. Following, we applied the design to dynamically document blood and lymphatic sprouting and patterning of the initially
avascular tadpole fin. Additionally, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin
had been designed for dynamic evaluation of loss-of-operate and acquire-of-perform phenotypes employing pharmacologic or genetic manipulation.
Together with angiography and lymphangiography to assess performance, Tg(Flk1:eGFP) reporter tadpoles conveniently allowed in depth lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP) tadpoles symbolize a functional design for functional lymph/angiogenomics and drug screening.
Creator contributions A.N., W.V., P.C. and M.D. developed experiments and wrote the manuscript A.N. and W.V. performed experiments and all quantification
P.H., I.G., A.D.-J., K.B., S.P. and T.L. assisted with experiments M.B. and A.L. carried out the FACS sorting S.V. assisted
with confocal microscopy P.A.K. presented important reagents.
Competing pursuits The authors have no competing passions to declare.
- Acquired March eleven, 2013.
- Acknowledged June 10, 2013.
- © 2013. Revealed by The Organization of Biologists Ltd
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