A transgenic Xenopus laevis reporter model to study lymphangiogenesis
The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene disc...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3773334/ https://www.ncbi.nlm.nih.gov/pubmed/24143274 http://dx.doi.org/10.1242/bio.20134739 |
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author | Ny, Annelii Vandevelde, Wouter Hohensinner, Philipp Beerens, Manu Geudens, Ilse Diez-Juan, Antonio Brepoels, Katleen Plaisance, Stéphane Krieg, Paul A. Langenberg, Tobias Vinckier, Stefan Luttun, Aernout Carmeliet, Peter Dewerchin, Mieke |
author_facet | Ny, Annelii Vandevelde, Wouter Hohensinner, Philipp Beerens, Manu Geudens, Ilse Diez-Juan, Antonio Brepoels, Katleen Plaisance, Stéphane Krieg, Paul A. Langenberg, Tobias Vinckier, Stefan Luttun, Aernout Carmeliet, Peter Dewerchin, Mieke |
author_sort | Ny, Annelii |
collection | PubMed |
description | The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the Xenopus tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent Xenopus reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic Tg(Flk1:eGFP) Xenopus laevis reporter lines expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality, Tg(Flk1:eGFP) reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP) tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening. |
format | Online Article Text |
id | pubmed-3773334 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | The Company of Biologists |
record_format | MEDLINE/PubMed |
spelling | pubmed-37733342013-10-18 A transgenic Xenopus laevis reporter model to study lymphangiogenesis Ny, Annelii Vandevelde, Wouter Hohensinner, Philipp Beerens, Manu Geudens, Ilse Diez-Juan, Antonio Brepoels, Katleen Plaisance, Stéphane Krieg, Paul A. Langenberg, Tobias Vinckier, Stefan Luttun, Aernout Carmeliet, Peter Dewerchin, Mieke Biol Open Research Article The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the Xenopus tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent Xenopus reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic Tg(Flk1:eGFP) Xenopus laevis reporter lines expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality, Tg(Flk1:eGFP) reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP) tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening. The Company of Biologists 2013-07-11 /pmc/articles/PMC3773334/ /pubmed/24143274 http://dx.doi.org/10.1242/bio.20134739 Text en © 2013. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Research Article Ny, Annelii Vandevelde, Wouter Hohensinner, Philipp Beerens, Manu Geudens, Ilse Diez-Juan, Antonio Brepoels, Katleen Plaisance, Stéphane Krieg, Paul A. Langenberg, Tobias Vinckier, Stefan Luttun, Aernout Carmeliet, Peter Dewerchin, Mieke A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title | A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title_full | A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title_fullStr | A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title_full_unstemmed | A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title_short | A transgenic Xenopus laevis reporter model to study lymphangiogenesis |
title_sort | transgenic xenopus laevis reporter model to study lymphangiogenesis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3773334/ https://www.ncbi.nlm.nih.gov/pubmed/24143274 http://dx.doi.org/10.1242/bio.20134739 |
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