June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Altered fate of corneal epithelial cells in Dstncorn1 mice
Author Affiliations & Notes
  • Yuyun Zhu
    Department of Medical Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Sharolyn Kawakami-Schulz
    Department of Medical Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Wei-Hua Lee
    Department of Medical Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Akihiro Ikeda
    Department of Medical Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Winston W Y Kao
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio, United States
  • Sakae Ikeda
    Department of Medical Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Yuyun Zhu, None; Sharolyn Kawakami-Schulz, None; Wei-Hua Lee, None; Akihiro Ikeda, None; Winston Kao, None; Sakae Ikeda, None
  • Footnotes
    Support  NIH R01 EY016108
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 966. doi:
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    • Get Citation

      Yuyun Zhu, Sharolyn Kawakami-Schulz, Wei-Hua Lee, Akihiro Ikeda, Winston W Y Kao, Sakae Ikeda; Altered fate of corneal epithelial cells in Dstncorn1 mice. Invest. Ophthalmol. Vis. Sci. 2017;58(8):966.

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

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Abstract

Purpose : Dstncorn1 mice, deficient mutants of the actin depolymerizing factor destrin, display epithelial hyperproliferation, inflammation and neovascularization in the cornea. In the Dstncorn1 corneal epithelium, we observed keratin 12 (Krt12)-positive cells that are also positive for keratin 15 (Krt15), a marker of corneal epithelial progenitor cells that normally reside in the limbus. The purpose of this study is to test the hypothesis that abnormal actin dynamics lead to transformation of Krt12-positive corneal epithelial cells into the less differentiated state in Dstncorn1 mice.

Methods : The mTmG double fluorescent cre reporter system was used to genetically label specific cell populations for cell lineage tracing. Specifically, we generated mice with the genotypes of Dstncorn1/corn1; Krt12rtTA/rtTA; tetO-cre; mTmG/+, and Dstncorn1/+; Krt12rtTA/rtTA; tetO-cre; mTmG/+. Doxycycline (dox) was administered through diet for 10 days to label the differentiated Krt12-positive cells with green fluorescence (mG), during which mT reporter gene cassette was deleted by cre expressed in Krt12-positive cells. The mG was chased upon the withdraw of dox from the diet for 0, 2 and 4 days, and excised corneas were subjected to immunohistochemistry using the Krt15 antibody. The corneal area positive for mG and the total corneal area were quantified using ImageJ. Analysis of variance was used with Tukey’s honest significant difference test for statistical analysis.

Results : The mG-positive area normalized to the total corneal area was the largest immediately after the completion of dox administration (day 0), and decreased in time both in control and Dstncorn1 mice. The mG-positive corneal area was less in Dstncorn1 mice compared to control mice at each time point. In the Dstncorn1cornea, we observed mG labeled cells that were also stained by Krt15 antibody.

Conclusions : Our findings indicate that the Krt12-positive terminally differentiated corneal epithelial cells transform to the less differentiated state in the corneal epithelium of Dstncorn1 mice. The mG-positive area was larger in control mice compared to Dstncorn1 mice at each time point, suggesting that cells in the Dstncorn1 corneal epithelium may have a higher renewal rate compared to control mice.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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