K12 Expression: Cornea-Type Epithelial Differentiation

Summary

Principal Investigator: WINSTON KAO
Abstract: Congenital and acquired corneal diseases are exemplified by altered genetic functions. To better treat corneal diseases, it is imperative to gain insights of gene functions during embryonic development and pathogenesis in adults. To determine the molecular and cellular mechanisms of corneal epithelial differentiation and wound healing, a mouse line expressing Cre recombinase by the corneal epithelium has been prepared by knock-in the phage Cre gene into the Krt12 allele via gene targeting techniques. Additional mouse lines using Cre/IoxP system are prepared from the Krt12Cre knock-in mice for corneal epithelium-specific expression of green fluorescent protein, conditional ablations of Jun and Fos in compound floxed transgenic mice, Krt12Cre/Cre-RosaSF, Krt12Cre/Cre-Junf/f (homozygous floxed Jun) and Krt12Cre/Cre-Fosf/f, respectively. Aim 1 is to determine differentiation and wound healing of corneal epithelium: Aim 1a is to examine the hypothesis that one of the two Krt12 alleles is preferentially used by individual corneal epithelial cells, Aim 1b is to determine the kinetics of differentiation during embryonic development and post natal growth of cornea, Aim 1c tests the hypothesis that corneal TAC (transit amplifying cells) cells of young mice are stem cell-like and of stem cell characteristics. To examine roles of AP-1 transcription factors on homeostasis of corneal epithelium and healing of corneal epithelium debridement, conditional Jun and Fos knockout mice (Krt12Cre/Cre-Junf/f and Krt12Cre/Cre-Fosf/f) are prepared. Aim 2 is to determine roles of Jun on cornea-type epithelium differentiation and wound healing. Corneas of adult Krt12Cre/Cre-Junf/f mice will be subject to immunohistochemistry, northern and in situ hybridization, to determine the roles of Jun on homeostasis of corneal epithelium. The AP-1 dimers will be determined by immune precipitation with anti-Fos antibodies and western blot with anti-Jun family member. The nuclear and cytosol proteins isolated from injured and un-injured Krt12Cre/Cre-Junf/f and Krt12+/+-Junf/f (control) corneas will be subject to proteomic analysis using 2-dimensional High Performance Liquid Electrophoresis (2D-HPLE). Aim 3 is to examine roles of Fos on corneal-type epithelium differentiation and corneal wound healing as described in Aim 2.
Funding Period: 1995-08-01 - 2010-05-31
more information: NIH RePORT

Top Publications

  1. ncbi Expression of keratin 12 and maturation of corneal epithelium during development and postnatal growth
    Noriko Tanifuji-Terai
    Department of Ophthalmology, University of Cincinnati College of Medicine, OH 45267 0527, USA
    Invest Ophthalmol Vis Sci 47:545-51. 2006
  2. ncbi Corneal morphogenesis during development and diseases
    Winston W Y Kao
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Eye Contact Lens 36:265-8. 2010
  3. pmc A pathogenic relationship between a regulator of the actin cytoskeleton and serum response factor
    Angela M Verdoni
    Department of Medical Genetics, University of Wisconsin, Madison, Wisconsin 53706, USA
    Genetics 186:147-57. 2010
  4. pmc Monoallelic expression of Krt12 gene during corneal-type epithelium differentiation of limbal stem cells
    Yasuhito Hayashi
    Department of Ophthalmology, University of Cincinnati, and Shriners Hospital for Children, Cincinnati, OH 45267 0838, USA
    Invest Ophthalmol Vis Sci 51:4562-8. 2010
  5. pmc Electrically assisted delivery of macromolecules into the corneal epithelium
    Jinsong Hao
    Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, 3225 Eden Ave, HPB 136, Cincinnati, OH 45267, USA
    Exp Eye Res 89:934-41. 2009
  6. pmc Gene delivery to cornea
    Jinsong Hao
    Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, Cincinnati, OH 45267, USA
    Brain Res Bull 81:256-61. 2010
  7. pmc A novel protocol of whole mount electro-immunofluorescence staining
    Hongshan Liu
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 15:505-17. 2009
  8. pmc Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice
    Daniel Y Weng
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 14:562-71. 2008
  9. pmc Excess FGF-7 in corneal epithelium causes corneal intraepithelial neoplasia in young mice and epithelium hyperplasia in adult mice
    Taiichiro Chikama
    Department of Ophthalmology, University of Cincinnati Medical Center, 3223 Eden Ave, Suite 350, Cincinnati, OH 45267 0527, USA
    Am J Pathol 172:638-49. 2008
  10. ncbi Signaling pathways in morphogenesis of cornea and eyelid
    Winston W Y Kao
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 6:9-23. 2008

Detail Information

Publications14

  1. ncbi Expression of keratin 12 and maturation of corneal epithelium during development and postnatal growth
    Noriko Tanifuji-Terai
    Department of Ophthalmology, University of Cincinnati College of Medicine, OH 45267 0527, USA
    Invest Ophthalmol Vis Sci 47:545-51. 2006
    ..To determine the kinetics of corneal epithelial maturation during embryonic development and postnatal growth...
  2. ncbi Corneal morphogenesis during development and diseases
    Winston W Y Kao
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Eye Contact Lens 36:265-8. 2010
    ..To review the use of genetically modified mouse lines for elucidating corneal morphogenesis during embryonic development and diseases...
  3. pmc A pathogenic relationship between a regulator of the actin cytoskeleton and serum response factor
    Angela M Verdoni
    Department of Medical Genetics, University of Wisconsin, Madison, Wisconsin 53706, USA
    Genetics 186:147-57. 2010
    ..Our study also demonstrates that Dstn is genetically upstream of Srf and defines a new functional role for SRF as the master regulator of a hyperproliferative, inflammatory phenotype accompanied by neovascularization...
  4. pmc Monoallelic expression of Krt12 gene during corneal-type epithelium differentiation of limbal stem cells
    Yasuhito Hayashi
    Department of Ophthalmology, University of Cincinnati, and Shriners Hospital for Children, Cincinnati, OH 45267 0838, USA
    Invest Ophthalmol Vis Sci 51:4562-8. 2010
    ..The purpose of this study was to characterize a Krt12-Cre knock-in mouse line for corneal epithelium-specific gene ablation and to analyze the allelic selection of the keratin 12 (Krt12) gene during corneal type-epithelium differentiation...
  5. pmc Electrically assisted delivery of macromolecules into the corneal epithelium
    Jinsong Hao
    Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, 3225 Eden Ave, HPB 136, Cincinnati, OH 45267, USA
    Exp Eye Res 89:934-41. 2009
    ..These results illustrate the feasibility of electrically assisted delivery of macromolecules such as siRNA into the cornea...
  6. pmc Gene delivery to cornea
    Jinsong Hao
    Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, Cincinnati, OH 45267, USA
    Brain Res Bull 81:256-61. 2010
    ..The combination of iontophoresis and electroporation was found to be effective in delivering siRNA but not plasmid DNA into the corneal epithelium. Nanocarriers such as polymeric micelles are promising methods of corneal gene delivery...
  7. pmc A novel protocol of whole mount electro-immunofluorescence staining
    Hongshan Liu
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 15:505-17. 2009
    ..To develop a new method of whole mount immunostaining that improves the penetration of staining reagents into the cornea and decreases non-specific binding and background...
  8. pmc Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice
    Daniel Y Weng
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 14:562-71. 2008
    ..To examine whether promiscuous Cre/LoxP recombination happens during gametogenesis in double transgenic mice carrying LoxP modified alleles and Cre transgene driven by tissue-specific promoter outside the gonads of adult mice...
  9. pmc Excess FGF-7 in corneal epithelium causes corneal intraepithelial neoplasia in young mice and epithelium hyperplasia in adult mice
    Taiichiro Chikama
    Department of Ophthalmology, University of Cincinnati Medical Center, 3223 Eden Ave, Suite 350, Cincinnati, OH 45267 0527, USA
    Am J Pathol 172:638-49. 2008
    ..Taken together, Krt12-rtTA/tet-O-FGF-7 mice may be a suitable animal model for the study of the molecular and cellular mechanisms of human OSSN...
  10. ncbi Signaling pathways in morphogenesis of cornea and eyelid
    Winston W Y Kao
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 6:9-23. 2008
    ..This review discusses known signaling transduction pathways involved in the morphogenesis of the cornea and eyelids during embryonic development and homeostasis in adults, using genetically modified experimental animal models...
  11. ncbi Eye drop delivery of nano-polymeric micelle formulated genes with cornea-specific promoters
    Yaw Chong Tong
    College of Pharmacy, Taipei Medical University, Taipei, Taiwan
    J Gene Med 9:956-66. 2007
    ..keratin 12 (K12) and keratocan (Kera3.2) promoters, by non-ionic poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) polymeric micelles (PM) to mouse and rabbit eyes, and investigates the underlying mechanisms...
  12. ncbi Transient receptor potential vanilloid 1 activation induces inflammatory cytokine release in corneal epithelium through MAPK signaling
    Fan Zhang
    Biological Sciences, The State University of New York, College of Optometry, New York, New York 10036, USA
    J Cell Physiol 213:730-9. 2007
    ..Taken together, TRPV1 receptors may play a role in mediating corneal epithelial inflammatory mediator secretion and subsequent hyperalgesia...
  13. ncbi Ocular surface tissue morphogenesis in normal and disease states revealed by genetically modified mice
    Winston W Y Kao
    Department of Ophthalmology and Cell Biology, University of Cincinnati, 3225 Eden Avenue, Cincinnati, OH 45267, USA
    Cornea 25:S7-S19. 2006
    ..These mouse lines can also be used as models for development of therapeutic treatment regimens of ocular surface diseases using gene therapy and stem cell strategies...
  14. pmc Changes in corneal basal epithelial phenotypes in an altered basement membrane
    I Jong Wang
    Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
    PLoS ONE 6:e14537. 2011
    ..To examine the corneal epithelial phenotype in an altered basement membrane...