Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaResults: Treatment of corneas with pellets containing thecombination of FGF2/LYVE-1 resulted in the outgrowth oflymphatic vessels with a mean neovascularization area of 0.12 ± 0.07mm 2 (n=20) whereas corneas with pellets containing FGF2 aloneshowed a mean neovascularization area of 0.23 ± 0.11 mm 2 (n=21) (p= 0.0008). Contrary no significant difference was found betweenVEGF-C induced lymphangiogenesis and the combination of VEGF-C/LYVE-1 induced lymphangiogenesis in the micropocket assay.Conclusions: LYVE-1 inhibited the FGF2-induced outgrowth oflymphatic vessels in a corneal micropocket assay suggesting thatLYVE-1 may bind FGF2 and interact with bFGF influencinglymphangiogenesis in vivo.Commercial Relationships: Birgit Regenfuss, None; NataliaPlatonova, None; Géraldine Miquel, None; Said Taouji, None;Eric Chevet, None; Andreas Bikfalvi, None; Claus Cursiefen,Gene Signal (C), Alcon (R), Allergan (R), Bayer (R)Support: German Research Foundation (DFG) SFB 643/B-10 andCu 47/4-1) to C.C. Agence National de la Recherche (ANR), InstitutNational du Cancer (INCA), Conseil Régional d’Aquitaine to A.B.and INCA, Association de Recherche contre le Cancer, Inserm(Avenir) to E.C.Program Number: 2101 Poster Board Number: D0240Presentation Time: 11:00 AM - 12:45 PMAnalysis of VEGFR-1 Isoforms in Pax6-deficient Mice CorneasPhillip A. Moore 1 , Peter J. Accola 1 , Barbara Artelt 1 , Megan Aarnio 1 ,James D. Lauderdale 2 . 1 Dept Small Animal Med/Surg, College VetMed Univ Georgia, Athens, GA; 2 Department of Cellular Biology,Franklin College of Arts and Sciences, The University of Georgia,Athens, GA.Purpose: To identify VEGFR-1 isoforms in the cornea of Pax6-deficient mice.Methods: The limbal region, central cornea and conjunctiva of Pax6+/- mice were compared to wild type mice for the presence ofVEGFR-1 and sVEGFR-1. Immunostaining was performed to wildtype (n=5) and Pax6 +/- mice (n= 5) with three antibodies: oneantibody to the N-terminus that detected both membrane and solubleVEGFR-1; one antibody to the C-terminus that detected themembrane bound form of VEGFR-1; and one antibody thatspecifically detected the soluble form of VEGFR-1. Immunoblottingwas performed on protein extracts prepared separately from thecornea and conjunctiva of wild type and Pax6 +/- mice.Results: Immunostaining revealed that both VEGFR-1 and sVEGFR-1 are present in the cornea of both wild type and Pax6 +/- mice.Immunoblotting revealed that both VEGFR-1 and sVEGFR-1 arepresent in the conjunctiva and central cornea of wild type and Pax6+/- mice. Interestingly, this analysis also revealed VEGFR-1 isoformsthat appeared to be specific to Pax6 +/- corneas.Conclusions: Our results are in contradiction to previous reports thatsVEGFR-1 is deficient in Pax6 +/- corneas. The presence ofsVEGFR-1 and VEGFR-1 isoforms specific to Pax6 +/- corneassuggests that a deficiency in sVEGFR-1 is not the cause for cornealneovascularization in Pax6 +/- mice.Commercial Relationships: Phillip A. Moore, None; Peter J.Accola, None; Barbara Artelt, None; Megan Aarnio, None; JamesD. Lauderdale, NoneSupport: UGA Veterinary Ophthalmology Research Fund, SharonStewart Aniridia Research Trust, Vision for Tomorrow FoundationProgram Number: 2102 Poster Board Number: D0241Presentation Time: 11:00 AM - 12:45 PMUse of Amaranthus leucocarpus lectin to determine corneal neovascularizationValeria L. Coria 1, 2 , Gibran A. Estua 1 , Alfredo Domínguez 1 , EdgarZenteno 2 , Jessica Nieves-Hernández 1 , Yonathan Garfias 1, 2 .1 Research Unit, Institute of Ophthalmology, Mexico city, Mexico;2 Biochemistry Department, Faculty of Medicine, UniversidadNacional Autónoma de México, Mexico city, Mexico.Purpose: The cornea is the interphase between the eye and theexternal environment, and is also the most powerful refractive surfacein the eye. In non-pathological conditions, the cornea is one of thefew avascular tissues from the body. The mechanism responsible forestablishing the avascular nature of the cornea is unknown; but, inseveral pathologycal situations like trauma, infection or surgicalprocedures, the stroma can be invaded by abnormal vessels(neovascularization) leading it to opacification. Theneovascularization is the creation of new vessels from preexistingones; in these abnormal vessels, the glycosylation could be aberrant.The glycosylation process, is the post translational modification inwhich saccharide residues get added to amino acid chains. Some ofthe tools used for studying neo-vessels in eye are lectins, such asGriffonia simplicifolia. Amaranthus leucocarpus (ALL) is a lectinthat recognizes specifically Galbeta1,3GalNAc carbohydratesstructures.The aim of this study was at determining Galbeta1,3GalNAcstructures in a corneal neovascularization murine modelMethods: This study was carried out in accordance with theInstitutional Animal Care and Use Committe and Vision Researchwith the ARVO statement for the Use of Animals in Ophthalmic andVision Research. All experiments were performed on 6-8 week-oldfemale BALB/c mice. Corneas were chemically burned with NaOH.Burned and healthy mice corneas were obtained, and were processedfor immunohistochemistry assays. ALL coupled to FITC was used toperform immunolabeling on corneal tissue; CD31 was used to colocatethe neovessel labeling of ALL-recognized structuresResults: The presence of ALL was exclusively observed in thechemically-burned mice corneal tissues, in contrast to the burnedtissues, there was not ALL-immunostaining in the healthy micecorneal tissues. Interestingly, ALL colocalized with CD31immunostaining in the burned and neovascularized corneal tissue.Conclusions: Amaranthus leucocarpus lectin is able to identifycorneal neo-vessels, suggesting that o-glycosylation process could bean important process in developing this aberrant corneal condition.Commercial Relationships: Valeria L. Coria, None; Gibran A.Estua, None; Alfredo Domínguez, None; Edgar Zenteno, None;Jessica Nieves-Hernández, Institute of Ophthalmology (P);Yonathan Garfias, Institute of Ophthalmology (P)Support: CONACYT 160286 and 167438Program Number: 2103 Poster Board Number: D0242Presentation Time: 11:00 AM - 12:45 PMNetrin-4 mediates corneal neovascularizationAnna-Karina Maier, Sabrina V. Klein, Norbert Kociok, AlineRiechardt, Enken Gundlach, Claudia Steger, Olaf Strauss, AntoniaM. Joussen. Charité, University Medicine Berlin, Berlin, Germany.Purpose: Netrin-4, a secreted protein, regulates neuronal andvascular growth. It is found in the basement membranes of bloodvessels. Previous results are contradictory supporting Netrin-4 aseither a pro- or anti-angiogenic factor. In the presented study weinvestigated the role of Netrin-4 in the mouse-model of sutureinduced,inflammatory corneal neovascularization.Methods: Corneal neovascularization was induced in Netrin-4-deficient (Ntn4-/-) and wild-type mice by suturing three 11-0 nylonintrastromally into the cornea. Vascularized area of cornealflatmounts were analyzed 14 days after suturing byimmunocytochemistry in conjunction withimage and statistical©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.