Abstract
Purpose:
The cornea is the interphase between the eye and the external environment, and is also the most powerful refractive surface in the eye. In non-pathological conditions, the cornea is one of the few avascular tissues from the body. The mechanism responsible for establishing the avascular nature of the cornea is unknown; but, in several pathologycal situations like trauma, infection or surgical procedures, the stroma can be invaded by abnormal vessels (neovascularization) leading it to opacification. The neovascularization is the creation of new vessels from preexisting ones; in these abnormal vessels, the glycosylation could be aberrant. The glycosylation process, is the post translational modification in which saccharide residues get added to amino acid chains. Some of the tools used for studying neo-vessels in eye are lectins, such as Griffonia simplicifolia. Amaranthus leucocarpus (ALL) is a lectin that recognizes specifically Galbeta1,3GalNAc carbohydrates structures. The aim of this study was at determining Galbeta1,3GalNAc structures in a corneal neovascularization murine model
Methods:
This study was carried out in accordance with the Institutional Animal Care and Use Committe and Vision Research with the ARVO statement for the Use of Animals in Ophthalmic and Vision Research. All experiments were performed on 6-8 week-old female BALB/c mice. Corneas were chemically burned with NaOH. Burned and healthy mice corneas were obtained, and were processed for immunohistochemistry assays. ALL coupled to FITC was used to perform immunolabeling on corneal tissue; CD31 was used to co-locate the neovessel labeling of ALL-recognized structures
Results:
The presence of ALL was exclusively observed in the chemically-burned mice corneal tissues, in contrast to the burned tissues, there was not ALL-immunostaining in the healthy mice corneal tissues. Interestingly, ALL colocalized with CD31 immunostaining in the burned and neovascularized corneal tissue.
Conclusions:
Amaranthus leucocarpus lectin is able to identify corneal neo-vessels, suggesting that o-glycosylation process could be an important process in developing this aberrant corneal condition.
Keywords: 554 immunohistochemistry