Transgenic Research

, Volume 21, Issue 3, pp 683–689 | Cite as

Transgenic mouse model expressing tdTomato under involucrin promoter as a tool for analysis of epidermal differentiation and wound healing

  • Petr Kasparek
  • Pavel Krenek
  • Halka Buryova
  • Sarka Suchanova
  • Inken Maria Beck
  • Radislav Sedlacek
Technical Report

Abstract

The epidermis is a stratified tissue composed of different keratinocyte layers that create a barrier protecting the body from external influences, pathogens, and dehydration. The barrier function is mainly achieved by its outermost layer, the stratum corneum. To create a mouse model to study pathophysiological processes in the outermost layers of the epidermis in vivo and in vitro we prepared a construct containing red fluorescent td-Tomato reporter sequence under the control of involucrin promoter and its first intron. Transgenic mice were generated by pronuclear injection and the expression and regulation of the transgene was determined by in vivo imaging and fluorescent microscopy. The promoter targeted the transgene efficiently and specifically into the outermost epidermal layers although weak expression was also found in epithelia of tongue and bladder. The regulation of expression in the epidermis, i.e. fluorescence intensity of the reporter, could be easily followed during wound healing and dermatitis. Thus, these transgenic mice carrying the tdTomato reporter could be used as a valuable tool to study impact of various genes dysregulating the epidermal barrier and to follow effects of therapeutic agents for treatment of skin diseases in vivo.

Keywords

Epidermis Involucrin tdTomato Wound healing Dermatitis Transgenic 

References

  1. Asamoto M, Fukushima S, Horike H, Tatemoto Y, Mori M (1989) Involucrin expression in urinary bladder carcinoma. Urol Res 17(5):279–283PubMedGoogle Scholar
  2. Banks-Schlegel S, Green H (1981) Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. J Cell Biol 90(3):732–737PubMedCrossRefGoogle Scholar
  3. Candi E, Schmidt R, Melino G (2005) The cornified envelope: a model of cell death in the skin. Nat Rev Mol Cell Biol 6(4):328–340. doi:10.1038/nrm1619 PubMedCrossRefGoogle Scholar
  4. Carroll JM, Taichman LB (1992) Characterization of the human involucrin promoter using a transient beta-galactosidase assay. J Cell Sci 103(Pt 4):925–930PubMedGoogle Scholar
  5. Carroll JM, Albers KM, Garlick JA, Harrington R, Taichman LB (1993) Tissue- and stratum-specific expression of the human involucrin promoter in transgenic mice. Proc Natl Acad Sci USA 90(21):10270–10274PubMedCrossRefGoogle Scholar
  6. Eckert RL, Green H (1986) Structure and evolution of the human involucrin gene. Cell 46(4):583–589PubMedCrossRefGoogle Scholar
  7. Elias PM (2005) Stratum corneum defensive functions: an integrated view. J Invest Dermatol 125(2):183–200PubMedGoogle Scholar
  8. Fuchs E, Green H (1981) Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell 25(3):617–625PubMedCrossRefGoogle Scholar
  9. Ghazizadeh S, Carroll JM, Taichman LB (1997) Repression of retrovirus-mediated transgene expression by interferons: implications for gene therapy. J Virol 71(12):9163–9169PubMedGoogle Scholar
  10. Ghazizadeh S, Doumeng C, Taichman LB (2002) Durable and stratum-specific gene expression in epidermis. Gene Ther 9(19):1278–1285. doi:10.1038/sj.gt.3301800 PubMedCrossRefGoogle Scholar
  11. Goebeler M, Gutwald J, Roth J, Sorg C (1991) The severity of irritant contact dermatitis in various strains of mice correlates with endothelial expression of migration inhibitory factor (MIF). Arch Dermatol Res 283(4):246–250PubMedCrossRefGoogle Scholar
  12. Ishida-Yamamoto A, Iizuka H (1995) Differences in involucrin immunolabeling within cornified cell envelopes in normal and psoriatic epidermis. J Invest Dermatol 104(3):391–395PubMedCrossRefGoogle Scholar
  13. Ishida-Yamamoto A, Iizuka H (1998) Structural organization of cornified cell envelopes and alterations in inherited skin disorders. Exp Dermatol 7(1):1–10PubMedCrossRefGoogle Scholar
  14. Kalinin AE, Kajava AV, Steinert PM (2002) Epithelial barrier function: assembly and structural features of the cornified cell envelope. BioEssays News Rev Mol Cell Dev Biol 24(9):789–800. doi:10.1002/bies.10144 CrossRefGoogle Scholar
  15. Kopan R, Traska G, Fuchs E (1987) Retinoids as important regulators of terminal differentiation: examining keratin expression in individual epidermal cells at various stages of keratinization. J Cell Biol 105(1):427–440PubMedCrossRefGoogle Scholar
  16. Magnaldo T, Bernerd F, Asselineau D, Darmon M (1992) Expression of loricrin is negatively controlled by retinoic acid in human epidermis reconstructed in vitro. Differ Res Biol Divers 49(1):39–46CrossRefGoogle Scholar
  17. Mannik J, Alzayady K, Ghazizadeh S (2010) Regeneration of multilineage skin epithelia by differentiated keratinocytes. J Invest Dermatol 130(2):388–397. doi:10.1038/jid.2009.244 PubMedCrossRefGoogle Scholar
  18. Mehrel T, Hohl D, Rothnagel JA, Longley MA, Bundman D, Cheng C, Lichti U, Bisher ME, Steven AC, Steinert PM et al (1990) Identification of a major keratinocyte cell envelope protein, loricrin. Cell 61(6):1103–1112PubMedCrossRefGoogle Scholar
  19. Merritt AJ, Berika MY, Zhai W, Kirk SE, Ji B, Hardman MJ, Garrod DR (2002) Suprabasal desmoglein 3 expression in the epidermis of transgenic mice results in hyperproliferation and abnormal differentiation. Mol Cell Biol 22(16):5846–5858PubMedCrossRefGoogle Scholar
  20. Nemes Z, Steinert PM (1999) Bricks and mortar of the epidermal barrier. Exp Mol Med 31(1):5–19PubMedGoogle Scholar
  21. Neufang G, Furstenberger G, Heidt M, Marks F, Muller-Decker K (2001) Abnormal differentiation of epidermis in transgenic mice constitutively expressing cyclooxygenase-2 in skin. Proc Natl Acad Sci USA 98(13):7629–7634. doi:10.1073/pnas.121574098 PubMedCrossRefGoogle Scholar
  22. Rice RH, Green H (1977) The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein. Cell 11(2):417–422PubMedCrossRefGoogle Scholar
  23. Rice RH, Green H (1979) Presence in human epidermal cells of a soluble protein precursor of the cross-linked envelope: activation of the cross-linking by calcium ions. Cell 18(3):681–694PubMedCrossRefGoogle Scholar
  24. Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22(12):1567–1572. doi:nbt1037[pii]10.1038/nbt1037 PubMedCrossRefGoogle Scholar
  25. Steinert PM, Marekov LN (1999) Initiation of assembly of the cell envelope barrier structure of stratified squamous epithelia. Mol Biol Cell 10(12):4247–4261PubMedGoogle Scholar
  26. Sumitomo S, Kumasa S, Iwai Y, Mori M (1986) Involucrin expression in epithelial tumors of oral and pharyngeal mucosa and skin. Oral Surg Oral Med Oral Pathol 62(2):155–163PubMedCrossRefGoogle Scholar
  27. Tubaro A, Dri P, Melato M, Mulas G, Bianchi P, Del Negro P, Della Loggia R (1986) In the croton oil ear test the effects of non steroidal antiinflammatory drug (NSAIDs) are dependent on the dose of the irritant. Agents Actions 19(5–6):371–373PubMedCrossRefGoogle Scholar
  28. Turksen K, Troy TC (2002) Permeability barrier dysfunction in transgenic mice overexpressing claudin 6. Development 129(7):1775–1784PubMedGoogle Scholar
  29. Walts AE, Said JW, Siegel MB, Banks-Schlegel S (1985) Involucrin, a marker of squamous and urothelial differentiation. An immunohistochemical study on its distribution in normal and neoplastic tissues. J Pathol 145(4):329–340. doi:10.1002/path.1711450406 PubMedCrossRefGoogle Scholar
  30. Wang X, Zinkel S, Polonsky K, Fuchs E (1997) Transgenic studies with a keratin promoter-driven growth hormone transgene: prospects for gene therapy. Proc Natl Acad Sci USA 94(1):219–226PubMedCrossRefGoogle Scholar
  31. Werner S, Weinberg W, Liao X, Peters KG, Blessing M, Yuspa SH, Weiner RL, Williams LT (1993) Targeted expression of a dominant-negative FGF receptor mutant in the epidermis of transgenic mice reveals a role of FGF in keratinocyte organization and differentiation. EMBO J 12(7):2635–2643PubMedGoogle Scholar
  32. Wilker E, Bol D, Kiguchi K, Rupp T, Beltran L, DiGiovanni J (1999) Enhancement of susceptibility to diverse skin tumor promoters by activation of the insulin-like growth factor-1 receptor in the epidermis of transgenic mice. Mol Carcinog 25(2):122–131PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Petr Kasparek
    • 1
  • Pavel Krenek
    • 1
    • 2
  • Halka Buryova
    • 1
  • Sarka Suchanova
    • 1
  • Inken Maria Beck
    • 1
    • 3
  • Radislav Sedlacek
    • 1
  1. 1.Department of Transgenic Models of DiseasesInstitute of Molecular Genetics of the ASCRPrague 4Czech Republic
  2. 2.Department of Cell Biology, Faculty of Science, Centre of the Region Hana for Biotechnological and Agricultural ResearchPalacky UniversityOlomoucCzech Republic
  3. 3.Institute of Biotechnology ASCRPrague 4Czech Republic

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