title: Limited versus total epithelial debridement ocular surface injury: Live fluorescence imaging of hemangiogenesis and lymphangiogenesis in Prox1-GFP/Flk1::Myr-mCherry mice.
abstract: Immunohistochemical staining experiments have shown that both hemangiogenesis and lymphangiogenesis occur following severe corneal and conjunctival injury and that the neovascularization of the cornea often has severe visual consequences. To better understand how hemangiogenesis and lymphangiogenesis are induced by different degrees of ocular injury, we investigated patterns of injury-induced corneal neovascularization in live Prox1-GFP/Flk1::myr-mCherry mice, in which blood and lymphatic vessels can be imaged simultaneously in vivo.
  
  
  
  The eyes of Prox1-GFP/Flk1::myr-mCherry mice were injured according to four models based on epithelial debridement of the: A) central cornea (a 1.5-mm-diameter circle of tissue over the corneal apex), B) total cornea, C) bulbar conjunctiva, and D) cornea+bulbar conjunctiva. Corneal blood and lymphatic vessels were imaged on days 0, 3, 7, and 10 post-injury, and the percentages of the cornea containing blood and lymphatic vessels were calculated.
  
  
  
  Neither central corneal nor bulbar conjunctival debridement resulted in significant vessel growth in the mouse cornea, whereas total corneal and corneal+bulbar conjunctival debridement did. On day 10 in the central cornea, total cornea, bulbar conjunctiva, and corneal+bulbar conjunctival epithelial debridement models, the percentage of the corneal surface that was occupied by blood vessels (hemangiogenesis) was 1.9±0.8%, 7.14±2.4%, 2.29±1%, and 15.05±2.14%, respectively, and the percentage of the corneal surface that was occupied by lymphatic vessels (lymphangiogenesis) was 2.45±1.51%, 4.85±0.95%, 2.95±1.27%, and 4.15±3.85%, respectively.
  
  
  
  Substantial corneal debridement was required to induce corneal neovascularization in the mouse cornea, and the corneal epithelium may therefore be partially responsible for maintaining corneal avascularity.
  
  
  
  Our study demonstrates that GFP/Flk1::myr-mCherry mice are a useful model for studying coordinated hemangiogenic and lymphangiogenic responses.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA. Electronic address: changr@uic.edu.
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Chang
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Ilham
authors.initials: I
authors.lastname: Putra
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Yu-Hui
authors.initials: YH
authors.lastname: Huang
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Chang
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Kyuyeon
authors.initials: K
authors.lastname: Han
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhong
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Xinbo
authors.initials: X
authors.lastname: Gao
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Shuangyong
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Dugas-Ford
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Tara
authors.initials: T
authors.lastname: Nguyen
authors.affiliation: Department of Surgery, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
authors.forename: Young-Kwon
authors.initials: YK
authors.lastname: Hong
authors.affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA. Electronic address: dazar@uic.edu.
authors.forename: Dimitri T
authors.initials: DT
authors.lastname: Azar
mesh_terms: D000818:Animals; D003315:Cornea / Q000254:growth & development* / Q000378:metabolism / Q000473:pathology; D065306:Corneal Injuries / Q000235:genetics* / Q000473:pathology; D016510:Corneal Neovascularization / Q000235:genetics* / Q000473:pathology; D003646:Debridement; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000254:growth & development / Q000473:pathology; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D042583:Lymphangiogenesis / Q000235:genetics; D042601:Lymphatic Vessels / Q000378:metabolism / Q000473:pathology; D051379:Mice; D025521:Tumor Suppressor Proteins / Q000235:genetics*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics*; D000098551:Prospero-Related Homeobox 1 Protein
chemical_list: D018398:Homeodomain Proteins; D025521:Tumor Suppressor Proteins; D000098551:Prospero-Related Homeobox 1 Protein; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Bulbar conjunctiva; Cornea; Corneal neovascularization (NV); Hemangiogenesis (HA); In vivo imaging; Lymphangiogenesis (LA)