July 2018
Volume 59, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2018
Endogenous Sonic hedgehog involvement in corneal epithelial cell proliferation in response to injury in mice
Author Affiliations & Notes
  • Kana Ichikawa
    Wakayama Medical University, Wakayama, Japan
  • Yuka Okada
    Wakayama Medical University, Wakayama, Japan
  • Osamu Yamanaka
    Wakayama Medical University, Wakayama, Japan
  • Chia-Yang Liu
    Optometry, Indiana University, Bloomington, Indiana, United States
  • Winston W Y Kao
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio, United States
  • Shizuya Saika
    Wakayama Medical University, Wakayama, Japan
  • Footnotes
    Commercial Relationships   Kana Ichikawa, None; Yuka Okada, None; Osamu Yamanaka, None; Chia-Yang Liu, None; Winston Kao, None; Shizuya Saika, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4419. doi:https://doi.org/
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      Kana Ichikawa, Yuka Okada, Osamu Yamanaka, Chia-Yang Liu, Winston W Y Kao, Shizuya Saika; Endogenous Sonic hedgehog involvement in corneal epithelial cell proliferation in response to injury in mice. Invest. Ophthalmol. Vis. Sci. 2018;59(9):4419. doi: https://doi.org/.

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

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Abstract

Purpose : A corneal epithelium-specific Sonic hedgehog (Shh) knockout mouse line is produced to examine the roles of endogenous Shh in epithelial wound healing. Its function in maintaining tissue homeostasis in adults remains elusive, although exogenous Shh enhances corneal epithelium repair (Saika S et al. IOVS, 2004).

Methods : Mice carrying Krt12-Cre (Hayashi Y et al. IOVS, 2010) and those carrying Shhf/f (Dassule H.R. et al. Development, 2000) were mated to generate a bi-transgenic Krt12Cre/wt/Shhf/f (ShhΔCE) mouse line. An epithelial debridement (2 mm in diameter) was produced at the center of demarcated cornea of right eye of 8-week-old male ShhΔCE and Shhf/wt (Krt12wt/wt/Shhf/wt) mice with a micro-surgical blade. Epithelial wound healing and cell proliferation were assessed at 6, 12, 18, 24 and 36 hrs post-injury by Image J software and BrdU immunostaining, respectively.

Results : Corneal epithelial debridement caused an up-regulation of cell proliferation in both ShhΔCE and Shhf/wt mice in comparison to naïve/uninjured corneas. The increment of BrdU-positive cells was counteracted in ShhΔCE mice compared with Shhf/wt in healing epithelium at 18 hrs after injury. The size of the remaining epithelial defect was not significantly altered in ShhΔCE as compared with Shhf/wt mice.

Conclusions : Endogenous Shh is required for up-regulation of cell proliferation in healing corneal epithelium in the later phase. Further experiment is required to elucidate the effects of complete loss of Shh on the corneal epithelial wound healing using homozygous Krt12Cre/Cre/Shhf/f mice.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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