Archives of Dermatological Research

, Volume 279, Issue 5, pp 335–340

Effect of growth environment on spatial expression of involucrin by human epidermal keratinocytes

  • F. M. Watt
  • P. Boukamp
  • J. Hornung
  • N. E. Fusenig
Original Contributions


Involucrin, the major protein precursor of the cornified envelope, is expressed during terminal differentiation of human keratinocytes, both in vivo and in vitro. In epidermis, the onset of synthesis is several layers above the basal layer, but in stratified cultures of keratinocytes on tissue culture plastic involucrin synthesis begins in the first suprabasal layer. To investigate the reason for this premature expression, the distribution of involucrin was studied in epidermis from different body sites, in organotypic cultures and in transplants of keratinocytes onto nude mice. We found that premature expression was not associated with poor morphological differentiation, because involucrin synthesis began immediately above the basal layer even when distinct basal, spinous, granular and cornified layers were formed in organotypic cultures recombined with dermis. The site of involucrin expression in culture did not depend on the number of cornified layers present. The only conditions which resulted in an upward shift in the site of synthesis were in 3-week old transplants on nude mice. We conclude that the site of onset of involucrin synthesis is not determined by the degree of morphological differentiation of the tissue, and discuss other factors which may be involved.

Key words

Keratinocytes Involucrin Organo typic cultures Transplants 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Asselineau D, Bernard BA, Bailly C, Darmon M, Pruniéras M (1986) Human epidermis reconstructed by culture: is it “normal”? J Invest Dermatol 86:181–186Google Scholar
  2. 2.
    Banks-Schlegel S, Green H (1980) Formation of epidermis by serially cultivated human epidermal cells transplanted as an epithelium to athymic mice. Transplantation 29:308–313Google Scholar
  3. 3.
    Banks-Schlegel S, Green H (1981) Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. J Cell Biol 90:732–737Google Scholar
  4. 4.
    Bernard BA, Reano A, Darmon YM, Thivolet J (1986) Precocious appearance of involucrin and epidermal transglutaminase during differentiation of psoriatic skin. Br J Dermatol 114:279–283Google Scholar
  5. 5.
    Boukamp P, Fusenig NE (1984) The influence of substrate and mesenchyme on the growth and differentiation of human kertinocytes in vitro. Eur J Cell Biol 33:4Google Scholar
  6. 6.
    Boukamp P, Rupniak HTR, Fusenig NE (1985) Environmental modulation of the expression of differentiation and malignancy in six human squamous cell carcinoma cell lines. Cancer Res 45:5582–5592Google Scholar
  7. 7.
    Breitkreutz D, Bohnert A, Herzmann E, Bowden PE, Boukamp P, Fusenig NE (1984) Differentiation specific functions in cultured and transplanted mouse keratinocytes: environmental influences on ultrastructure and keratin expression. Differentiation 26:154–169Google Scholar
  8. 8.
    Dover R, Potten CS (1983) Cell cycle kinetics of cultured human epidermal keratinocytes. J Invest Dermatol 80: 423–429Google Scholar
  9. 9.
    Fusenig NE (1986) Mammalian epidermal cells in culture. In: Bereiter-Hahn J Matoltsy AG, Richards KS (eds) Biology of the integument, vol 2, Vertebrates. Springer, Berlin Heidelberg New York, pp 409–442Google Scholar
  10. 10.
    Fusenig NE, Breitkreutz D, Lueder M, Boukamp P, Worst PKM (1981) Keratinization and structural organization in epidermal cell cultures. In: Schweiger HG (ed) International cell biology 1980–1981. Springer, Berlin Heidelberg New York, pp 1004–1014Google Scholar
  11. 11.
    Fusenig NE, Breitkreutz D, Dzarlieva RT, Boukamp P, Bohnert A, Tilgen W (1983) Growth and differentiation characteristics of transformed keratinocytes from mouse and human skin in vitro and in vivo. J Invest Dermatol 81:168s-175sGoogle Scholar
  12. 12.
    Green H (1977) Terminal differentiation of cultured human epidermal cells. Cell 11:405–416Google Scholar
  13. 13.
    Green H (1980) The keratinocyte as differentiated cell type. Harvey Lect 74:101–139Google Scholar
  14. 14.
    Green H, Watt FM (1982) Regulation by vitamin A of envelope cross-linking in cultured keratinocytes derived from different human epithelia. Mol Cell Biol 2:1115–1117Google Scholar
  15. 15.
    Green H, Fuchs E, Watt F (1982) Differentiated structural components of the keratinocyte. Cold Spring Harbor Sympos Quant Biol 46:293–301Google Scholar
  16. 16.
    Holbrook KA, Hennings H (1983) Phenotypic expression of epidermal cells in vitro: a review. J Invest Dermatol 81:11s-24sGoogle Scholar
  17. 17.
    Karasek MA (1983) Culture of human keratinocytes in liquid medium. J Invest Dermatol 81:24s-28sGoogle Scholar
  18. 18.
    Mackenzie IC, Fusenig NE (1983) Regeneration of organized epithelial structure. J Invest Dermatol 81:189s-194sGoogle Scholar
  19. 19.
    Murphy GF, Flynn TC, Rice RH, Pinkus GS (1984) Involucrin expression in normal and neoplastic human skin: a marker for keratinocyte differentiation. J Invest Dermatol 82:453–457Google Scholar
  20. 20.
    Potten CS (1983) Kinetic organisation in squamous epithelium. In: Wright NA, Camplejohn RS (eds) Psoriasis: cell proliferation. Churchill Livingstone, Edinburgh, pp 149–162Google Scholar
  21. 21.
    Pruniéras M, Régnier M, Woodley D (1983) Methods for cultivation of keratinocytes with an air-liquid interface. J Invest Dermatol 81:28s-33sGoogle Scholar
  22. 22.
    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 18:681–694Google Scholar
  23. 23.
    Simon M, Green H (1984) Participation of membrane-associated proteins in the formation of the cross-linked envelope of the keratinocyte. Cell 36:827–834Google Scholar
  24. 24.
    Sun TT, Green H (1976) Differentiation of the epidermal keratinocyte in cell culture: formation of the cornified envelope. Cell 9:511–521Google Scholar
  25. 25.
    Watt FM (1983) Involucrin and other markers of keratinocyte terminal differentiation. J Invest Dermatol 81:100s-103sGoogle Scholar
  26. 26.
    Watt FM (1984) Selective migration of terminally differentiating cells from the basal layer of cultured human epidermis. J Cell Biol 98:16–21Google Scholar
  27. 27.
    Watt FM, Green H (1981) Involucrin synthesis is correlated with cell size in human epidermal cultures. J Cell Biol 90:738–742Google Scholar
  28. 28.
    Watt FM, Green H (1982) Stratification and terminal differentiation of cultured epidermal cells. Nature 295:434–436Google Scholar
  29. 29.
    Watt FM, Mattey DL, Garrod DR (1984) Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol 99:2211–2215Google Scholar
  30. 30.
    Weiss RA, Eichner R, Sun TT (1984) Monoclonal antibody analysis of keratin expression in epidermal diseases: a 48-and 56-kDalton keratin as molecular markers for hyperproliferative keratinocytes. J Cell Biol 98:1397–1406Google Scholar
  31. 31.
    Worst PKM, Mackenzie IC, Fusenig NE (1982) Reformation of organized epidermal structure by transplantation of suspensions and cultures of epidermal and dermal cells. Cell Tissue Res 225:65–77Google Scholar
  32. 32.
    Wu YJ, Parker LM, Binder NE, Beckett MA, Sinard JH, Griffiths CT, Rheinwald JG (1982) The mesothelial keratins: a new family of cytoskeletal proteins identified in cultured mesothelial cells and nonkeratinizing epithelia. Cell 31:693–703Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • F. M. Watt
    • 1
  • P. Boukamp
    • 2
  • J. Hornung
    • 2
  • N. E. Fusenig
    • 2
  1. 1.Molecular Cell Biology LaboratoryKennedy Institute of RheumatologyLondonEngland
  2. 2.Division of Differentiation and Carcinogenesis in vitro, Institute of BiochemistryDeutsches Krebsforschungszentrum (DKFZ)HeidelbergFRG
  3. 3.Imperial Cancer Research FundLondonEngland

Personalised recommendations