Abstract
Although VEGFR-3 deficiency disrupts blood vascular development during early embryogenesis, the underlying mechanism was not clear. To characterize its function in angiogenesis and lymphangiogenesis, we employed two genetically modified mouse models in this study, targeting the coding region for the ligand-binding domain (Vegfr3ΔLBD) or the tyrosine kinase domain with an inactivation point mutation (Vegfr3TKmut). We show that lymphatic growth was disrupted in Vegfr3ΔLBD/ΔLBD and Vegfr3TKmut/TKmut mice, but blood vessels developed normally in both embryo and yolk sac. Interestingly, in Vegfr3ΔLBD/ΔLBD but not Vegfr3TKmut/TKmut mice, lymph sac was present but there was lack of lymphangiogenic sprouting. We further demonstrate that both the wild-type and mutant forms of VEGFR-3 could form heterodimers with VEGFR-2, and decreased the level of phospho-VEGFR-2 and the downstream phospho-Erk1/2 in endothelial cells when they were treated with VEGF-A. These findings indicate that signaling mediated via VEGFR-3 activation by its cognate ligands (VEGF-C/-D) is not required for angiogenesis, and that VEGFR-3 may play a role in this process by modulating VEGFR-2-mediated signals.
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Acknowledgements
We thank Dr Lena Claesson-Welsh (Uppsala University), and PIs of Model Animal Research Center (MARC, Nanjing University) for the helpful discussion about the work, and Yanlan Cao, Wenting Shi and all the staff in the MARC Animal facility of Nanjing University for excellent technical assistance. This work was financially supported by grants from the National Natural Science Foundation of China (30771069, 30671038, and 30930028), the Ministry of Science and Technology of China (2006CB943500), and the Ministry of Education of China (NCET: Program for New Century Excellent Talents in University).
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( Supplementary information is linked to the online version of the paper on Cell Research website.)
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Supplementary information, Figure S1
VEGFR-3 targeting strategy and vascular analysis in somite regions. (PDF 202 kb)
Supplementary information, Figure S2
Analysis of VEGFR-3 expression in blood vessels. (PDF 690 kb)
Supplementary information, Figure S3
Analysis of receptor autophosphorylation of three different forms of VEGFR-3 (VEGFR-3WT, VEGFR-3Tkmut, and VEGFR-3ΔLBD). (PDF 104 kb)
Supplementary information, Figure S4
Analysis of somite vascular growth in Vegfr3TKmut/TKmut mice. (PDF 150 kb)
Supplementary information, Figure S5
Analysis of retina and tumor angiogenesis in Vegfr3WT/TKmut mice. (PDF 233 kb)
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Zhang, L., Zhou, F., Han, W. et al. VEGFR-3 ligand-binding and kinase activity are required for lymphangiogenesis but not for angiogenesis. Cell Res 20, 1319–1331 (2010). https://doi.org/10.1038/cr.2010.116
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DOI: https://doi.org/10.1038/cr.2010.116
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