title: Comparison of EphA receptor tyrosine kinases and ephrinA ligand expression to EphB-ephrinB in vascularized corneas.
abstract: OBJECTIVE
  Eph cell surface receptors and their ligands, ephrins, are involved in neuronal patterning and neovascularization. Our purpose is to compare and characterize the expression of ephrinA ligands and EphA receptors to ephrinB ligands and EphB receptors in excised mouse corneal tissue, in corneal epithelial and keratocyte cell lines, and during corneal angiogenesis.
  
  
  METHODS
  Mouse corneal epithelial cells and keratocytes were immortalized using SV40T antigen viral infection of primary cultures. The immortalized epithelial cells and keratocytes were cloned and characterized using antibodies to keratin, vimentin, integrin alpha5beta1, and alpha-smooth muscle actin. Basic fibroblast growth factor pellets were implanted to induce corneal neovascularization. The eyes of wild-type, ephrinB2(tlacZ/+), and EphB4(tlacZ/+) heterozygous mice were harvested and sectioned 7 days after pellet implantation. Confocal immunohistochemistry was performed to compare the expression of the Eph/ephrinA family (EphA1-8, ephrinA1-5) and Eph/ephrinB family (EphB1-4, EphB6 ephrinB1-3).
  
  
  RESULTS
  EphA1, EphA3, ephrinA1, ephrinA2, EphB1, EphB4, ephrinB1, and ephrinB2 were detected in wild-type mouse corneal epithelial cells and keratocytes. EphA2 was immunolocalized only in epithelial cells. Also, EphA3, ephrinA1, EphB1, EphB4, and ephrinB1 were immunolocalized to the corneal epithelium and stroma. In the vascularized corneas, ephrinB1 was immunolocalized mainly to the keratocytes around the vessels, and ephrinB2, EphB1, and EphB4 were colocalized mainly with CD31 to the vascular endothelial cells.
  
  
  CONCLUSIONS
  The characterization of ephrin ligand and Eph receptor expression during cornea angiogensis in this study suggests that the Eph/ephrin family of receptor tyrosine kinases and their ligands may play a role in the regulation of corneal angiogenesis.
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois, Chicago, IL 60612, USA.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kojima
authors.forename: Tae-Young
authors.initials: TY
authors.lastname: Chung
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Chang
authors.forename: Rony
authors.initials: R
authors.lastname: Sayegh
authors.forename: Fabio H
authors.initials: FH
authors.lastname: Casanova
authors.forename: Dimitri T
authors.initials: DT
authors.lastname: Azar
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D016510:Corneal Neovascularization / Q000378:metabolism*; D003319:Corneal Stroma / Q000166:cytology; D036342:Ephrins / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D039081:Integrin alpha5beta1 / Q000378:metabolism; D007633:Keratins / Q000378:metabolism; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D036081:Receptors, Eph Family / Q000378:metabolism*; D014746:Vimentin / Q000378:metabolism
chemical_list: D000199:Actins; D036342:Ephrins; D039081:Integrin alpha5beta1; D008024:Ligands; D014746:Vimentin; D007633:Keratins; D036081:Receptors, Eph Family