June 2013
Volume 54, Issue 15
ARVO Annual Meeting Abstract  |   June 2013
Transglutaminase-2 Knockdown Effect Epithelial-Mesenchymal Transition during Corneal Wound Healing
Author Affiliations & Notes
  • Aihua Hou
    Singapore Eye Research Institute, Singapore, Singapore
  • Louis Tong
    Singapore Eye Research Institute, Singapore, Singapore
    Singapore National Eye Center, Singapore, Singapore
  • Footnotes
    Commercial Relationships Aihua Hou, None; Louis Tong, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 3872. doi:
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      Aihua Hou, Louis Tong; Transglutaminase-2 Knockdown Effect Epithelial-Mesenchymal Transition during Corneal Wound Healing. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3872.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: Transglutaminase-2 (TGM-2) is a multifunctional cross-linking enzyme involved in wound healing. Cornea wound closure was delayed in TGM-2-/- knockout mice. Also, Knockdown of TGM-2 reduced epithelial cell adhesion and migration. Epithelial-mesenchymal transition (EMT) was suggested to be involved in wound healing in cornea epithelial cells. This study aimed to investigate whether TGM2 defect perturbs EMT during corneal wound healing.

Methods: The central cornea of anesthetized mice was marked by a trephine, and the epithelium was peeled off using forceps under a dissecting microscope. Mouse eyes were harvest at different time points and embedded in OCT, and then were sectioned with cryostat at 8µM thickness. Immunofluorescent staining was performed on mouse eye sections using anti-Slug, anti-Twist and anti-β-Catenin antibodies.

Results: In TGM2+/+ mice, EMT marker Slug was observed 4hrs following corneal abrasion, while in TGM2-/- mice, slug was observed at 24hours. Interestingly, in both TGM2+/+ and TGM2-/- mice, Slug was expressed only in the nuclei of cells at basal layer of the impaired epithelium. By immunostaining, at both naive and wounded cornea epithelium, TGM2 +/+ mice had stronger β-catenin expression than TGM2-/- mice. In TGM2+/+ mice, strong β-Catenin staining was observed at the advancing edge of corneal epithelium 4 hrs after abrasion, this phenomenon was absent at the wound edge in TGM2-/- mice. Weak Twist staining was observed in both naive and wounded corneal epithelium, but no obvious difference between TGM2+/+ and TGM2-/- mice.

Conclusions: TGM-2 knockdown delayed Slug expression in wounded corneal epithelium, and also reduced β-Catenin expression. These findings may suggest perturbation of EMT, which has implications for wound response. The therapeutic significance is in the modulation of wound healing via TGM-2 in ocular surface diseases.

Keywords: 482 cornea: epithelium • 765 wound healing • 512 EMT (epithelial mesenchymal transition)  

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