Cell surface glycoconjugate abnormalities and corneal epithelial wound healing in the pax6+/- mouse model of aniridia-related keratopathy.
OBJECTIVE
Congenital aniridia due to heterozygosity for Pax6 is associated with ocular surface disease, including keratopathy. This study investigated how defects in glycoconjugate component of the cell surface of Pax6+/- could cause the abnormal cellular migration phenotypes associated with the disease.


METHODS
Immunohistochemistry, lectin-based histochemistry, conventional staining techniques, and proteomic assays were performed on eyes and cultured corneal epithelial cells from wild-type and Pax6+/- littermates. Wild-type cells were manipulated in culture to replicate the glycoconjugate abnormalities found in Pax6 heterozygotes and determine the consequences for wound healing.


RESULTS
Multiple glycoconjugate defects were found in Pax6-mutant cells. Lectin cytochemistry of corneal epithelial cells suggested a partial failure of glycoprotein trafficking. Blocking cell surface carbohydrate moieties in wild-type corneal cells caused wound-healing delays similar to those seen in untreated Pax6+/- cells.


CONCLUSIONS
Alterations to the cell surface glycoconjugate signature of Pax6+/- corneal epithelia restrict the ability of cells to initiate migration in response to wounding. This underlies the observed wound-healing delay in cultured Pax6+/- epithelia.
Kucerova
Romana
R
School of Medical Sciences, University of Aberdeen, Institute of Medical Sciences, Aberdeen, United Kingdom.
Ou
Jingxing
J
Lawson
Diane
D
Leiper
Lucy J
LJ
Collinson
J Martin
JM
D000818:Animals; D015783:Aniridia / Q000378:metabolism*; D001692:Biological Transport; D018929:Cell Culture Techniques; D003316:Corneal Diseases / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000378:metabolism*; D005136:Eye Proteins / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D006001:Glycoconjugates / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000502:physiology*; D007150:Immunohistochemistry; D037102:Lectins; D008562:Membrane Glycoproteins / Q000172:deficiency*; D051379:Mice; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000502:physiology*; D020543:Proteome / Q000378:metabolism; D012097:Repressor Proteins / Q000502:physiology*; D014945:Wound Healing
D005136:Eye Proteins; D006001:Glycoconjugates; D018398:Homeodomain Proteins; D037102:Lectins; D008562:Membrane Glycoproteins; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D020543:Proteome; D012097:Repressor Proteins