Cell and Tissue Research

, Volume 267, Issue 2, pp 347–356 | Cite as

Expression of vimentin by rabbit corneal epithelial cells during wound repair

  • N. SundarRaj
  • J. D. Rizzo
  • S. C. Anderson
  • J. P. Gesiotto


Intermediate filaments of epithelial cells generally consist of specific combinations of keratins. However, cultured epithelial cells from certain tissues and some epithelial tumors have been shown also to express vimentin. In the present study, the expression of vimentin by epithelial cells in healing corneal wounds (partial thickness penetrating wounds) and in tissue culture was analyzed. Both immunohistochemical and immunotransblot analyses indicated that although vimentin was not detected in the normal rabbit corneal epithelium in vivo, cultured rabbit corneal epithelial cells co-express keratins and vimentin. At 1 day post-wounding, vimentin was not detectable in the epithelial cells that had covered the denuded stroma. However, at 2 days post-wounding, the epithelium at the base of the epithelial plug immunoreacted with both anti-vimentin and antikeratin monoclonal antibodies. Immunotransblot analyses of the extracts of the epithelial plugs confirmed the presence of vimentin (Mr=58k). The 58k band was not detected in the extract of normal rabbit corneal epithelium. At day/5, vimentin was no longer detectable in the epithelium. This study demonstrated that corneal epithelial cells transiently co-express vimentin and keratins in vivo during wound healing and in tissue culture. The time-course of the transient expression of vimentin suggests that the vimentin expression in the epithelial cells during healing is not linked to cell proliferation or to the centripetal migration of the epithelium during early stages (first 24 h) of healing, but may be linked to cell-matrix interactions or the migration of basal cells in the upward direction at the following stage of healing.

Key words

Cornea Epithelium Wound healing Keratin Vimentin Rabbit 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Azumi N, Battifora H (1987) The distribution of vimentin and keratin in epithelial and nonepithelial neoplasms. A comprehensive immunohistochemical study on formalin-and alcohol-fixed tumors. Am J Clin Pathol 88:286–296Google Scholar
  2. Ben-Ze'ev A (1984) Differential control of cytokeratins and vimentin synthesis by cell-cell contact and cell spreading in cultured epithelial cells. J Cell Biol 99:1424–1433Google Scholar
  3. Ben-Ze'ev A (1985) Cell-cell interaction and cell configuration related control of cytokeratins and vimentin expression in epithelial cells and in fibroblasts. Ann N Y Acad Sci 455:597–613Google Scholar
  4. Ben-Ze'ev A (1986) Tumor promoted-induced disruption of junctional complexes in cultured corneal epithelial cells is followed by inhibition of cytokeratin and desmoplakin synthesis. Exp Cell Res 161:335–352Google Scholar
  5. Binder PS, Wickham MG, Zavala EY, Akers PH (1980) Corneal anatomy and wound healing. In: Symposium on medical and surgical diseases of the cornea. Transactions of the New Orleans Academy of Ophthalmology. Mosby, St. Louis, pp 1–35Google Scholar
  6. Boyer B, Tucker GC, Valles AM, Franke WW, Thiery JP (1989) Rearrangements of desmosomal and cytoskeletal proteins during the transition from epithelial to fibroblastoid organization in cultured rat bladder carcinoma cells. J Cell Biol 109:1495–1509Google Scholar
  7. Cabral F, Willingham MC, Gottesman MM (1980) Ultrastructural localization of 10 nm filaments of an insoluble 58K protein in cultured fibroblasts. J Histochem Cytochem 28:653–662Google Scholar
  8. Caselitz J, Becker J, Seifert G, Weber K Osborn M (1984) Coexpression of keratin and vimentin filaments in adenoid cystic carcinomas of salivary glands. Virchows Arch [A] 403:337–344Google Scholar
  9. Connell ND, Rheinwald JG (1983) Regulation of the cytoskeleton in mesothelial cells: reversible loss of keratin and increase in vimentin during rapid growth in culture. Cell 34:245–253Google Scholar
  10. Dulbecco R, Allen R, Okada S, Bowman M (1983) Functional changes of intermediate filaments in fibroblastic cells revealed by a monoclonal antibody. Proc Natl Acad Sci USA 80:1915–1918Google Scholar
  11. Elder JH, Munson M (1984) Modifications of immunoblotting technique for detection and quantitation of infectious virus in cell monolayers. Biotechniques, pp 170–172Google Scholar
  12. Fellini SA, Bennett GS, Toyama Y, Holtzer H (1978) Biochemical and immunological heterogeneity of 100 Å filament subunits from different chick cell types. Differentiation 12:59–69Google Scholar
  13. Franke WW, Schmid E, Weber K (1978) Different intermediatesized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci USA 75:5034–5038Google Scholar
  14. Franke WW, Mayer D, Schmid E, Denk H, Borenfreund E (1981) Differences of expression of cytoskeletal proteins in cultured rat hepatocytes and hepatoma cells. Exp Cell Res 134:345–365Google Scholar
  15. Geiger B, Kreis TE, Gigi O, Schmid E, Mittnacht S, Jorcano JL, von Bassewitz DB, Franke WW (1984) Dynamic rearrangements of cytokeratins in living cells. In: Levine AJ, Woude GFV, Topp WC, Watson JD (eds) Cancer Cells. The Transformed Phenotype. Cold Spring Harbor Laboratories, Cold Spring Harbor, NY, pp 201–214Google Scholar
  16. Goodman WM, SundarRaj N, Garone M, Arffa RC, Thoft RA (1989) Unique parameters in the healing of linear partial thickness penetrating corneal incisions in rabbit: immunohistochemical evaluation. Curr Eye Res 8:305–316Google Scholar
  17. Grone HJ, Weber K, Grone E, Helmchen U, Osborn M (1987) Coexpression of keratin and vimentin in damaged and regenerating tubular epithelia of the kidney. Am J Pathol 129:1–8Google Scholar
  18. Hanna C (1966) Proliferation and migration of epithelial cells during corneal wound rapair in the rabbit and the rat. Am J Ophthalmol 61:55–62Google Scholar
  19. Jackson BW, Grund C, Schmid E, Burki K, Franke WW, Illmensee K (1980) Formation of cytoskeletal elements during mouse embryogenesis. Differentiation 17:161–189Google Scholar
  20. Jumblatt MM, Neufeld AH (1983) B-adrenergic and serotonergic responsiveness of rabbit corneal epithelial cells in culture. Invest Ophthalmol Vis Sci 24:1139–1143Google Scholar
  21. Kartenbeck J, Schwechheimer K, Moll R, Franke WW (1984) Attachment of vimentin filaments to desmosomal plaques in human meningiomal cells and arachnoidal tissue. J Cell Biol 98:1072–1081Google Scholar
  22. Kitano S, Goldman JN (1966) Cytologic and histochemical changes in corneal wound repair. Arch Ophthal 76:345–354Google Scholar
  23. Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  24. Lane EB, Goodman SL, Trejdosiewicz LK (1982) Disruption of the keratin filament network during epithelial cell division. EMBO J 1:1365–1372Google Scholar
  25. Lane EB, Hogan BLM, Kurkinen M, Garrels JJ (1983) Co-expression of vimentin and cytokeratins in parietal endoderm cells of early mouse embryo. Nature 303:701–704Google Scholar
  26. LaRocca PJ, Rheinwald JG (1984) Coexpression of simple epithelial keratins and vimentin by human mesothelium and mesothelioma in vivo and in culture. Cancer Res 44:2991–2999Google Scholar
  27. Lemp MA (1976) Cornea and sclera. Arch Ophthalmol 94:473–490Google Scholar
  28. Leong AS, Gilham P, Milios J (1988) Cytokeratin and vimentin intermediate filament proteins in benign and neoplastic prostatic epithelium. Histopathology 13:435–442Google Scholar
  29. Lowry OH, Rosenbrough AL, Farr AL, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:690–697Google Scholar
  30. Mahrle G, Bolling R, Osborn M, Weber K (1983) Intermediate filaments of the vimentin and prekeratin type in human epidermis. J Invest Dermatol 81:46–48Google Scholar
  31. McLean IW, Nakane DK (1974) Periodate-lysine-paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083Google Scholar
  32. Moll R, Franke WW, Schiller D, Geiger B, Krepler R (1982) The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31:11–24Google Scholar
  33. Moll R, Krepler R, Franke WW (1983) Complex cytokeratin polypeptide patterns observed in certain human carcinomas. Differentiation 23:256–269Google Scholar
  34. Osborn M (1985) Summary: intermediate filaments. Ann N Y Acad Sci 455:669–681Google Scholar
  35. Risen LA, Binder PS, Nayak SK (1987) Intermediate filaments and their organization in human corneal endothelium. Invest Ophthalmol Vis Sci 28:1933–1938Google Scholar
  36. Robb RM, Kuwabara T (1964) Corneal wound healing. II An autoradiographic study of the cellular components. Arch Ophthalmol 72:401–408Google Scholar
  37. Rodrigues MM, Rowden G, Hackett J, Bakos I (1981) Langerhans cells in the normal conjunctiva and peripheral corneal selected species. Invest Ophthalmol Vis Sci 21:759–765Google Scholar
  38. Santini D, Bazzocchi F, Paladini G, Gelli MC, Ricci M, Mazzoleni G, Martinelli G (1987) Intermediate-sized filaments proteins (keratin, vimentin, desmin) in metaplastic carcinomas, carcinosarcomas and stromal sarcomas of the breast. Int J Biol Markers 2:83–86Google Scholar
  39. Schermer A, Galvin S, Sun T-T (1986) Differentiation related expression of a major 64K corneal keratin in vivo and in culture suggest limbal location of corneal epithelial cells in the limbal epithelium. J Cell Biol 103:49–62Google Scholar
  40. Schmid E, Franke WW, Grund C, Schiller DL, Kolb H, Paweletz N (1983a) An epithelial cell line with elongated myoid morphology derived from bovine mammary gland. Exp Cell Res 146:309–328Google Scholar
  41. Schmid E, Schiller DL, Grund C, Stadler J, Franke WW (1983b) Tissue type-specific expression of intermediate filament proteins in a cultured epithelial cell line from bovine mammary gland. J Cell Biol 96:37–50Google Scholar
  42. Smelser GK, Ozanics V (1972) Reaction of the cornea to injury and wound healing. In: Symposium on the cornea, Transactions of the New Orleans Academy of Ophthalmology. Mosby, St. Louis, pp 121–131Google Scholar
  43. Steinert PM, Liem RK (1990) Intermediate filament dynamics. Cell 60:521–523Google Scholar
  44. Steinert PM, Roop DR (1988) Molecular and cellular biology of intermediate filaments. Annu Rev Biochem 57:593–625Google Scholar
  45. Stoesser TR, Church RL, Brown SI (1978) Partial characterization of human collagen and procollagen secreted by human corneal fibroblasts in culture. Invest Ophthalmol Vis Sci 17:264–271Google Scholar
  46. SundarRaj N, Martin J, SundarRaj CV (1983) Cell-surface associated proteins of corneal fibroblasts: dissection with monoclonal antibodies. J Cell Biochem 21:277–287Google Scholar
  47. SundarRaj N, Willson J, Gregory JD, Damle SP (1985) Monoclonal antibodies to proteokeratan sulfate or rabbit corneal stroma. Curr Eye Res 4:49–54Google Scholar
  48. SundarRaj N, Anderson S, Barbacci-Tobin E (1988) An intermediate filament-associated developmentally regulated protein in corneal fibroblasts. Curr Eye Res 7:937–946Google Scholar
  49. SundarRaj N, Geiss MJ, Fantes F, Hanna K, Anderson SC, Thompson KP, Thoft RA, Waring GO (1990) Healing of excimer laser ablated monkey corneas: an immunohistochemical evaluation. Arch Ophthalmol 108:1604–1610Google Scholar
  50. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354Google Scholar
  51. Viale G, Gambacorta M, Dell'Orto P, Coggi G (1988) Coexpression of cytokeratins and vimentin in common epithelial tumours of the ovary: an immunocytochemical study of eighty-three cases. Virchows Arch [A] 413:91–101Google Scholar
  52. Virtanen I, Lehto VP, Lehtonen E, Vartio T, Stenman S, Kurki P, Wagner O, Small JV, Dahl D, Bradley RA (1981) Expression of intermediate filaments in cultured cells. J Cell Sci 50:45–63Google Scholar
  53. Williamson JSP, DiMarco S, Streilein JW (1987) Immunobiology of Langerhans cells on the ocular surface. Invest Ophthalmol Vis Sci 28:1527–1532Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • N. SundarRaj
    • 1
  • J. D. Rizzo
    • 1
    • 2
  • S. C. Anderson
    • 1
    • 2
  • J. P. Gesiotto
    • 1
    • 2
  1. 1.Department of OphthalmologyThe Eye Ear InstitutePittsburghUSA
  2. 2.The University of Pittsburgh School of MedicinePittsburghUSA

Personalised recommendations