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

Rodolfo A. Elizondo, Zhaohong Yin, Xiaowen Lu, Mitchell A. Watsky

Research output: Contribution to journalArticle

26 Scopus citations

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, Ca2+, 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.

Original languageEnglish (US)
Pages (from-to)5245-5251
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number8
DOIs
StatePublished - Jul 24 2014

Keywords

  • Corneal epithelium
  • Tight junctions
  • Vitamin D
  • Wound healing

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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