March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Allelic Expression of Tet-On rtTA Transgenic Driver Mouse Lines in Ocular Surface Tissues
Author Affiliations & Notes
  • Jianhua Zhang, Sr.
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Hongshan Liu
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Yong Yuan
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Yujin Zhang
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Chia-Yang Liu
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Mindy Call
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Minh-Thanh Nguyen
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Winston W. Kao
    Ophthalmology, University of Cincinnati, Cincinnati, Ohio
  • Footnotes
    Commercial Relationships  Jianhua Zhang, Sr., None; Hongshan Liu, None; Yong Yuan, None; Yujin Zhang, None; Chia-Yang Liu, None; Mindy Call, None; Minh-Thanh Nguyen, None; Winston W. Kao, None
  • Footnotes
    Support  NIH/NEI EY#13755, EY#011845, Research to Prevent Blindness, Ohio Lions Eye Research Foundation
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 4738. doi:
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    • Get Citation

      Jianhua Zhang, Sr., Hongshan Liu, Yong Yuan, Yujin Zhang, Chia-Yang Liu, Mindy Call, Minh-Thanh Nguyen, Winston W. Kao; Allelic Expression of Tet-On rtTA Transgenic Driver Mouse Lines in Ocular Surface Tissues. Invest. Ophthalmol. Vis. Sci. 2012;53(14):4738.

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

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Abstract

Purpose: : Understanding the expression pattern of tet-on driver mice can facilitate the interpretation of phenotypes following gene specific modification in vivo. The expression pattern of Pax6-rtTA (P6R), Krt12-rtTA (K12R), and Krt14-rtTA (K14R) transgenes were examined in the ocular surface epithelium and Kera-rtTA (KR) and Wnt1-Cre transgenes in stromal cells during ocular surface development.

Methods: : Triple transgenic P6R/tet-O-Cre (TC)/ROSAmTmG (mTG), K12R/TC/mTG, K14R/TC/mTG, KR/TC/mTG and double Wnt1-Cre/mTG were obtained by breeding and the expression patterns of rtTA and Wnt1-Cre transgenes were determined by EGFP expression following the expression of TC by doxycycline (Dox) induction and without Dox for Wnt1-Cre. Quantitative PCR was employed to determine the hetero- and homozygosity of rtTA in the driver mice.

Results: : In adult mice, both heterozygous K12R and K14R drivers showed mosaic and spiral EGFP expression patterns upon Dox induction, while homozygous K12R and K14R had a uniform pattern of EGFP expression. P6R drove EGFP expression in basal cells of the corneal epithelium and in lens epithelial and fiber cells in 3 week old mice that were induced at E0, E12.5 and E16.5. The EGFP+ cells gradually diminished as the mouse aged. Following pulse induction at E11, EGFP+ cells were first found in the trunk at E13 and then in the eye field at E14-E15 and postnatal P0. The number of EGFP+ cells was more abundant in homozygous P6R mice than in heterozygous mice. EGFP+ cells were found in all of the stromal cells of the eye of Wnt1-Cre/mTG mice, despite homo- or heterozygosity of the Wnt1-Cre transgene. In contrast the corneal endothelium showed a mosaic expression pattern, suggesting that multiple neural crest cell lineages may contribute to the corneal endothelium. In KR/TC/mTG mice, strong EGFP+ keratocytes were primarily located in the anterior stroma and weaker EGFP+ cells located at the posterior stroma, similar to that observed with Kera-Cre/mTG mice.

Conclusions: : Our observations of the EGFP expression pattern within the ocular surface using K12R and K14R driver mice are consistent with the notion that these transgenes are subjected to allelic regulation albeit the molecular mechanism remains unknown. The expression pattern of P6R suggests that epithelial stem cells do not utilize this transgene. Both Wnt1-Cre and KR are ubiquitously used by periocular mesenchymal cells.

Keywords: cornea: epithelium • development • cornea: storage 
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