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Simultaneous in vivo imaging of blood and lymphatic vessel growth in Prox1– GFP /Flk1::myr–mCherry mice
Author(s) -
Zhu Jimmy,
DugasFord Jennifer,
Chang Michael,
Purta Patryk,
Han KyuYeon,
Hong YoungKwon,
Dickinson Mary E.,
Rosenblatt Mark I.,
Chang JinHong,
Azar Dimitri T.
Publication year - 2015
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.13234
Subject(s) - lymphangiogenesis , angiogenesis , in vivo , lymphatic system , lymphatic vessel , green fluorescent protein , pathology , microbiology and biotechnology , vascular endothelial growth factor , chemistry , biology , cancer research , medicine , metastasis , vegf receptors , biochemistry , genetics , cancer , gene
The ability to visually observe angiogenesis and lymphangiogenesis simultaneously and repeatedly in living animals would greatly enhance our understanding of the inter‐dependence of these processes. To generate a mouse model that allows such visualization via in vivo fluorescence imaging, we crossed Prox1– GFP mice with Flk1::myr–mCherry mice to generate Prox1– GFP /Flk1::myr–mCherry mice, in which lymphatic vessels emit green fluorescence and blood vessels emit red fluorescence. Corneal neovascularization was induced in these mice using three injury models: implantation of a vascular endothelial growth factor ( VEGF ) pellet, implantation of a basic fibroblast growth factor (bFGF) pellet, and alkali burn injury. Vessel growth was observed in vivo by stereomicroscopy on days 0, 3, 7 and 10 after pellet implantation or alkali injury as well as in flat‐mounted corneas via confocal microscopy after the final in vivo imaging time point. We observed blood and lymphatic vessel growth in all three models, with the most significant growth occurring from days 0–7. Upon VEGF stimulation, the growth kinetics of blood and lymphatic vessels were similar. Blood vessels exhibited similar growth patterns in VEGF ‐ and bFGF‐stimulated corneas. Alkali burn injury induced robust angiogenesis and lymphangiogenesis. The intrinsic fluorescence of blood and lymphatic endothelial cells in Prox1– GFP /Flk1::myr–mCherry mice permitted simultaneous in vivo imaging of angiogenesis and lymphangiogenesis. This allowed us to differentiate the processes as well as observe their inter‐dependence, and will be valuable in development of therapies targeting angiogenesis and/or lymphangiogenesis.