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Invest Ophthalmol Vis Sci. 2014 Jul 24;55(8):5245-51. doi: 10.1167/iovs.13-13553.

Effect of vitamin D receptor knockout on cornea epithelium wound healing and tight junctions.

Author information

1
Department of Cellular Biology and Anatomy, Georgia Regents University, Augusta, Georgia, United States.
2
Department of Anatomy and Neuroscience, University of Tennessee Health Science Center, Memphis, Tennessee, United States.

Abstract

PURPOSE:

Our laboratory previously determined that vitamin D3, the vitamin D receptor (VDR), and 1α hydroxylase are present and active in the eye. In this study, we examined the effects of VDR knockout on wound healing, the tight junction-associated proteins occludin and ZO-1, and tight junction numbers in mouse corneas.

METHODS:

Epithelial wounds (2-mm) were made with an agar brush on 4-week-old and 10-week-old wild-type, heterozygous, and VDR knockout mouse corneas. Mice were on a normal or high lactose, Ca(2+), and PO₄(-) diet. Wound-healing area was measured over time. Real-time PCR was used to quantify occludin and ZO-1 message expression. Western blot was used for protein expression. Transmission electron microscopy was used to examine corneal epithelium and endothelium tight junctions. Immunofluorescence was used to examine epithelial ZO-1 distribution.

RESULTS:

Results showed a decreased healing rate in 10-week-old VDR knockout mice compared with wild-types. Vitamin D receptor knockout mice on the special diet had no difference in healing rate compared with wild-types. Real-time PCR showed decreased expression of occludin and ZO-1 in 10-week-old VDR knockout mice compared with wild-types. Western blot of 10-week-old knockout mouse corneas showed decreased occludin expression compared with wild-types. Transmission electron microscopy showed a significant difference in tight junction numbers in VDR knockouts versus wild-types. Immunofluorescence showed a change in ZO-1 distribution among genotypes.

CONCLUSIONS:

Vitamin D receptor knockout affects mouse corneal epithelium wound healing and tight junction integrity.

KEYWORDS:

corneal epithelium; tight junctions; vitamin D; wound healing

PMID:
25061117
PMCID:
PMC4142771
DOI:
10.1167/iovs.13-13553
[Indexed for MEDLINE]
Free PMC Article

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