May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Effects of cJun on Cytoplasmic Proteins Profile in Injured Corneal Epithelium
Author Affiliations & Notes
  • C. W. Kao
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • L. Zhou
    Singapore Eye Research Institute, Singapore, Singapore
  • H. Liu
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • R. W. Beuerman
    Ophthalmology,
    Singapore Eye Research Institute, Singapore, Singapore
  • W. W. Y. Kao
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Footnotes
    Commercial Relationships  C.W. Kao, None; L. Zhou, None; H. Liu, None; R.W. Beuerman, None; W.W.Y. Kao, None.
  • Footnotes
    Support  Grants from NIH EY010556, Research to Prevent Blindness, Ohio Lions Eye Research Foundation
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3379. doi:
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      C. W. Kao, L. Zhou, H. Liu, R. W. Beuerman, W. W. Y. Kao; Effects of cJun on Cytoplasmic Proteins Profile in Injured Corneal Epithelium. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3379.

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

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Abstract

Purpose: : Present studies examined the effects of cJun on the cytoplasmic proteins profile in injured corneal epithelium.

Methods: : cJun was conditionally ablated in corneal epithelium by feeding doxycycline for two weeks to triple transgenic Krt12rtTA/rtTA/tetO-Cre/cJunfloxed/floxed (cJunΔCE/ΔCE) mice. Corneal epithelium debridement (2 mm in diameter) was created in the center of corneas of experimental cJunΔCE/ΔCE (dox-induced) and cJunCEf/CEf (un-induced) triple transgenic mice. Cytoplasmic proteins isolated from injured epithelium healed for 6 and 12 h were subjected to proteomic analysis of multiplex quantitative proteomics method, iTRAQ (isobaric Tagging for Relative and Absolute protein Quantification), coupled with two dimensional nano-liquid chromatography (2D nanoLC) and Q-TOF tandem mass spectrometry (MS/MS), was used to simultaneously determine relative changes in the proteome of corneal epithelium.

Results: : About 150 proteins were identified by proteomic analysis. Data obtained from un-injured corneas, the ablation of cJun in triple transgenic cJunΔCE/ΔCE mice did not cause any significant changes (greater than 2 folds increase and decrease) in cytoplasmic proteins profile of corneal epithelium as compare to that of triple transgenic cJunCEf/CEf mice. There were little changes of cytoplasmic proteins profile in the injured corneal epithelium healed for 6 h in either cJunΔCE/ΔCE or cJunCEf/CEf mice. After 12 h of debridement, there were significant changes in proteins profile in corneas of both cJunΔCE/ΔCE and cJunCEf/CEf mice. Several proteins exhibited greater than five fold increase in comparison to the uninjured epithelium, serum albumin precursor, major urinary protein 4precursor, lacrimal androgen-binding proteins, prostatic steroid-binding protein, α1-antitrypsin precursor, hemopexin precursor, etc. The patterns of proteins profile changes were independent of the presence of cJun even though in many cases there were lower up-regulation of the proteins in the absence of cJun.

Conclusions: : cJun is not essential for maintaining the profile of cytoplasmic proteins in uninjured corneal epithelium. Epithelium debridement caused an up-regulation of several cytoplasmic proteins in 12 h. Absence of cJun did not abolish such up-regulation caused by injury. It is possible other members of may compensate the lost function of cJun in corneal epithelium of cJunΔCE/ΔCE mice.

Keywords: wound healing • transgenics/knock-outs • proteomics 
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