Longitudinal changes to tight junction expression and endothelial cell integrity in a mouse model of sterile corneal inflammation

Laura E. Downie, Janet Choi, Jeremiah K.H. Lim, Holly R. Chinnery

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Abstract

PURPOSE. We previously reported that applying toll-like receptor (TLR) ligands to an injured cornea induces corneal edema at 24 hours, which subsides by 1 week. We tested the hypotheses that endothelial expression of the tight-junction protein, zonula occludens-1 (ZO-1), would be altered during experimental sterile corneal inflammation and that endothelial cell density (ECD) would remain unaffected. METHODS. Anesthetized C57BL/6J mice received central 1-mm corneal abrasions followed by topical application of saline or cytosine-phosphate-guanosine oligodeoxynucleotide (CpGODN, TLR-9 agonist). At 24 hours, 1 week and 4 weeks post treatment, spectral-domain optical coherence tomography images were captured. Eyes were enucleated and processed for zonula occludens-1 (ZO-1) immunofluorescent staining. Corneal flatmounts were analyzed for endothelial ZO-1 expression, cell density, polymegethism, and polymorphism. Corneal stromal inflammatory cell infiltration was evaluated at 4 weeks by immunostaining for CD45. RESULTS. Central corneal thickness (CCT) was increased in CpG-ODN treated eyes at 24 hours, had normalized by 1 week, but was again thickened by 4 weeks. In eyes with CpG-ODN, endothelial cell ZO-1 expression was reduced at 24 hours but returned to normal levels by 1 week. Endothelial cell density was not altered at 24 hours or 1 week. By 4 weeks, only CpGODN eyes showed relatively reduced ECD, as well as large numbers of CD45+ cells in the stroma. Changes to ECD correlated with CCT (r = -0.53, P < 0.01). Compared with naïve controls, more saline-and CpG-ODN-treated eyes exhibited polymegethism. CONCLUSIONS. This study provides novel insights into the interplay between endothelial cell integrity, corneal edema, and chronic stromal leukocyte activation during sterile corneal inflammation in mice.

Original languageEnglish
Pages (from-to)3477-3484
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume57
Issue number7
DOIs
Publication statusPublished - Jun 2016
Externally publishedYes

Bibliographical note

This work is licensed under a Creative Commons Attribution 4.0 International License.

Keywords

  • Corneal endothelial cells
  • Inflammation
  • Toll-like receptor

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