title: Simultaneous in vivo imaging of blood and lymphatic vessel growth in Prox1-GFP/Flk1::myr-mCherry mice.
abstract: 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.
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Jimmy
authors: J
authors: Zhu
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Jennifer
authors: J
authors: Dugas-Ford
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Michael
authors: M
authors: Chang
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Patryk
authors: P
authors: Purta
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Kyu-Yeon
authors: KY
authors: Han
authors: Department of Surgery, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
authors: Young-Kwon
authors: YK
authors: Hong
authors: Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA.
authors: Mary E
authors: ME
authors: Dickinson
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Mark I
authors: MI
authors: Rosenblatt
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Jin-Hong
authors: JH
authors: Chang
authors: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA.
authors: Dimitri T
authors: DT
authors: Azar
mesh_terms: D000468:Alkalies / Q000633:toxicity; D000818:Animals; D002057:Burns, Chemical / Q000378:metabolism / Q000473:pathology; D065306:Corneal Injuries / Q000378:metabolism / Q000473:pathology*; D016510:Corneal Neovascularization; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D007124:Immunoenzyme Techniques; D008164:Luminescent Proteins / Q000378:metabolism*; D042583:Lymphangiogenesis; D042601:Lymphatic Vessels / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D025521:Tumor Suppressor Proteins / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000502:physiology*; D000097573:Red Fluorescent Protein; D000098551:Prospero-Related Homeobox 1 Protein
chemical_list: D000468:Alkalies; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D025521:Tumor Suppressor Proteins; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2; D000097573:Red Fluorescent Protein; D000098551:Prospero-Related Homeobox 1 Protein; C467480:vascular endothelial growth factor A, mouse
keywords: angiogenesis; cornea; endothelial cells; in vivo imaging; lymphangiogenesis