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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. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Present studies examined the effects of cJun on the cytoplasmic proteins profile in injured corneal epithelium.
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.
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.
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.
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