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Modulation of ocular surface glycocalyx barrier function by a galectin-3 N-terminal deletion mutant and membrane-anchored synthetic glycopolymers.
- Source :
-
PloS one [PLoS One] 2013 Aug 19; Vol. 8 (8), pp. e72304. Date of Electronic Publication: 2013 Aug 19 (Print Publication: 2013). - Publication Year :
- 2013
-
Abstract
- Background: Interaction of transmembrane mucins with the multivalent carbohydrate-binding protein galectin-3 is critical to maintaining the integrity of the ocular surface epithelial glycocalyx. This study aimed to determine whether disruption of galectin-3 multimerization and insertion of synthetic glycopolymers in the plasma membrane could be used to modulate glycocalyx barrier function in corneal epithelial cells.<br />Methodology/principal Findings: Abrogation of galectin-3 biosynthesis in multilayered cultures of human corneal epithelial cells using siRNA, and in galectin-3 null mice, resulted in significant loss of corneal barrier function, as indicated by increased permeability to the rose bengal diagnostic dye. Addition of β-lactose, a competitive carbohydrate inhibitor of galectin-3 binding activity, to the cell culture system, transiently disrupted barrier function. In these experiments, treatment with a dominant negative inhibitor of galectin-3 polymerization lacking the N-terminal domain, but not full-length galectin-3, prevented the recovery of barrier function to basal levels. As determined by fluorescence microscopy, both cellobiose- and lactose-containing glycopolymers incorporated into apical membranes of corneal epithelial cells, independently of the chain length distribution of the densely glycosylated, polymeric backbones. Membrane incorporation of cellobiose glycopolymers impaired barrier function in corneal epithelial cells, contrary to their lactose-containing counterparts, which bound to galectin-3 in pull-down assays.<br />Conclusions/significance: These results indicate that galectin-3 multimerization and surface recognition of lactosyl residues is required to maintain glycocalyx barrier function at the ocular surface. Transient modification of galectin-3 binding could be therapeutically used to enhance the efficiency of topical drug delivery.
- Subjects :
- Animals
Biological Transport
Cellobiose chemistry
Cells, Cultured
Cornea chemistry
Cornea cytology
Epithelial Cells chemistry
Galectin 3 deficiency
Glycocalyx chemistry
Glycoconjugates chemical synthesis
Glycoconjugates metabolism
Humans
Lactose chemistry
Mice
Mice, Knockout
Mucins chemistry
Mucins metabolism
Permeability
Protein Multimerization
Protein Structure, Tertiary
Rose Bengal metabolism
Structure-Activity Relationship
Cornea drug effects
Epithelial Cells drug effects
Galectin 3 genetics
Glycocalyx drug effects
Glycoconjugates pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 8
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 23977277
- Full Text :
- https://doi.org/10.1371/journal.pone.0072304