Tissue and cellular characterisation of nucleolin in a murine model of corneal angiogenesis.
OBJECTIVE
Corneal neovascularisation (CNV), with consequent loss of transparency, is due to an imbalance of proangiogenic factors. Cell-surface nucleolin (NCL) has been associated with neo-angiogenesis. There are studies identifying NCL translocation from nucleus to the cell surface, which is essential for endothelial cell proliferation. To find the possible role of NCL in the generation of corneal neovessels, the aim of this study is to characterise the NCL presence and cell-localisation in non-injured corneas, as well as to describe the changes in NCL cell and tissue localisation in CNV, and to analyse the effect of bevacizumab on NCL cellular and tissular distribution.


METHODS
Suture-induced CNV was performed in mice. The corneal tissues were obtained and the histological and co-immunofluorescence assays were performed using different proteins, such as CD31, cadherin and isolectin B4. To determine the possible role of VEGF in NCL presence and localisation in our CNV model, bevacizumab was concomitantly used.


RESULTS
Nucleolin was principally observed in the nucleus of the basal epithelial cells of normal corneas. Interestingly, angiogenesis-induced changes were observed in the localisation of NCL, not only in tissue but also at the cellular level where NCL was extranuclear in epithelial cells, stromal cells and neovessels. In contrast, these changes were reverted when bevacizumab was used. Besides, NCL was able to stain only aberrant corneal neovessels in comparison with retinal vessels.


CONCLUSIONS
NCL mobilisation outside the nucleus during angiogenesis could have a possible role as a proangiogenic molecule in the corneal tissue.
Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800, Mexico City, Mexico.|Faculty of Veterinary Medicine and Animal Husbandry, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico.
Joaquín
J
Quiroz-Mercado
Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800, Mexico City, Mexico.
Norma
N
Ramírez-Velázquez
Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800, Mexico City, Mexico.
Graciela
G
Partido
Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico.
Edgar
E
Zenteno
Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico.
Raúl
R
Chávez
Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico.
Concepción
C
Agundis-Mata
Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800, Mexico City, Mexico.|Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico.
Maria Carmen
MC
Jiménez-Martínez
Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800, Mexico City, Mexico. yogarfias@institutodeoftalmologia.org.|Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Avenida Universidad 3000, 04510, Mexico City, Mexico. yogarfias@institutodeoftalmologia.org.
Yonathan
http://orcid.org/0000-0003-3753-1035
Y
Garfias
D000818:Animals; D003315:Cornea / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D016510:Corneal Neovascularization / Q000175:diagnosis / Q000378:metabolism*; D004195:Disease Models, Animal; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins; D010750:Phosphoproteins / Q000096:biosynthesis*; D016601:RNA-Binding Proteins / Q000096:biosynthesis*; D011817:Rabbits; D000097796:Nucleolin
D009687:Nuclear Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin
Angiogenesis; Cornea; Nucleolin