Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Feb 1;96(2):494–509. doi: 10.1083/jcb.96.2.494

Morphology, behavior, and interaction of cultured epithelial cells after the antibody-induced disruption of keratin filament organization

PMCID: PMC2112303  PMID: 6187752

Abstract

The organization of intermediate filaments in cultured epithelial cells was rapidly and radically affected by intracellularly injected monoclonal antikeratin filament antibodies. Different antibodies had different effects, ranging from an apparent splaying apart of keratin filament bundles to the complete disruption of the keratin filament network. Antibodies were detectable within cells for more than four days after injection. The antibody-induced disruption of keratin filament organization had no light-microscopically discernible effect on microfilament or microtubule organization, cellular morphology, mitosis, the integrity of epithelial sheets, mitotic rate, or cellular reintegration after mitosis. Cell-to-cell adhesion junctions survived keratin filament disruption. However, antibody injected into a keratinocyte-derived cell line, rich in desmosomes, brought on a superfasciculation of keratin filament bundles, which appeared to pull desmosomal junctions together, suggesting that desmosomes can move in the plane of the plasma membrane and may only be 'fixed' by their anchoring to the cytoplasmic filament network. Our observations suggest that keratin filaments are not involved in the establishment or maintenance of cell shape in cultured cells.

Full Text

The Full Text of this article is available as a PDF (9.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albrecht-Buehler G. Daughter 3T3 cells. Are they mirror images of each other? J Cell Biol. 1977 Mar;72(3):595–603. doi: 10.1083/jcb.72.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderton B. H. Intermediate filaments: a family of homologous structures. J Muscle Res Cell Motil. 1981 Jun;2(2):141–166. doi: 10.1007/BF00711866. [DOI] [PubMed] [Google Scholar]
  3. Aubin J. E., Osborn M., Franke W. W., Weber K. Intermediate filaments of the vimentin-type and the cytokeratin-type are distributed differently during mitosis. Exp Cell Res. 1980 Sep;129(1):149–165. doi: 10.1016/0014-4827(80)90340-7. [DOI] [PubMed] [Google Scholar]
  4. Aubin J. E., Weber K., Osborn M. Analysis of actin and microfilament-associated proteins in the mitotic spindle and cleavage furrow of PtK2 cells by immunofluorescence microscopy. A critical note. Exp Cell Res. 1979 Nov;124(1):93–109. doi: 10.1016/0014-4827(79)90260-x. [DOI] [PubMed] [Google Scholar]
  5. Ball E. H., Singer S. J. Association of microtubules and intermediate filaments in normal fibroblasts and its disruption upon transformation by a temperature-sensitive mutant of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6986–6990. doi: 10.1073/pnas.78.11.6986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ben-Ze'ev A., Farmer S. R., Penman S. Mechanisms of regulating tubulin synthesis in cultured mammalian cells. Cell. 1979 Jun;17(2):319–325. doi: 10.1016/0092-8674(79)90157-0. [DOI] [PubMed] [Google Scholar]
  7. Blose S. H. Ten-nanometer filaments and mitosis: maintenance of structural continuity in dividing endothelial cells. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3372–3376. doi: 10.1073/pnas.76.7.3372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bray D. Surface movements during the growth of single explanted neurons. Proc Natl Acad Sci U S A. 1970 Apr;65(4):905–910. doi: 10.1073/pnas.65.4.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brûlet P., Babinet C., Kemler R., Jacob F. Monoclonal antibodies against trophectoderm-specific markers during mouse blastocyst formation. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4113–4117. doi: 10.1073/pnas.77.7.4113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dellagi K., Brouet J. C., Perreau J., Paulin D. Human monoclonal IgM with autoantibody activity against intermediate filaments. Proc Natl Acad Sci U S A. 1982 Jan;79(2):446–450. doi: 10.1073/pnas.79.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dembitzer H. M., Herz F., Schermer A., Wolley R. C., Koss L. G. Desmosome development in an in vitro model. J Cell Biol. 1980 Jun;85(3):695–702. doi: 10.1083/jcb.85.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Denk H., Franke W. W., Eckerstorfer R., Schmid E., Kerjaschki D. Formation and involution of Mallory bodies ("alcoholic hyalin") in murine and human liver revealed by immunofluorescence microscopy with antibodies to prekeratin. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4112–4116. doi: 10.1073/pnas.76.8.4112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Franke W. W., Schiller D. L., Moll R., Winter S., Schmid E., Engelbrecht I., Denk H., Krepler R., Platzer B. Diversity of cytokeratins. Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues. J Mol Biol. 1981 Dec 25;153(4):933–959. doi: 10.1016/0022-2836(81)90460-5. [DOI] [PubMed] [Google Scholar]
  15. Franke W. W., Schmid E., Breitkreutz D., Lüder M., Boukamp P., Fusenig N. E., Osborn M., Weber K. Simultaneous expression of two different types of intermediate sized filaments in mouse keratinocytes proliferating in vitro. Differentiation. 1979;14(1-2):35–50. doi: 10.1111/j.1432-0436.1979.tb01010.x. [DOI] [PubMed] [Google Scholar]
  16. Franke W. W., Schmid E., Grund C., Geiger B. Intermediate filament proteins in nonfilamentous structures: transient disintegration and inclusion of subunit proteins in granular aggregates. Cell. 1982 Aug;30(1):103–113. doi: 10.1016/0092-8674(82)90016-2. [DOI] [PubMed] [Google Scholar]
  17. Fuchs E. V., Coppock S. M., Green H., Cleveland D. W. Two distinct classes of keratin genes and their evolutionary significance. Cell. 1981 Nov;27(1 Pt 2):75–84. doi: 10.1016/0092-8674(81)90362-7. [DOI] [PubMed] [Google Scholar]
  18. Fuge H. Ultrastructure of the mitotic spindle. Int Rev Cytol Suppl. 1977;(6):1–58. [PubMed] [Google Scholar]
  19. Gawlitta W., Osborn M., Weber K. Coiling of intermediate filaments induced by microinjection of a vimentin-specific antibody does not interfere with locomotion and mitosis. Eur J Cell Biol. 1981 Dec;26(1):83–90. [PubMed] [Google Scholar]
  20. Geiger B., Singer S. J. Association of microtubules and intermediate filaments in chicken gizzard cells as detected by double immunofluorescence. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4769–4773. doi: 10.1073/pnas.77.8.4769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gordon W. E., 3rd, Bushnell A., Burridge K. Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. Cell. 1978 Feb;13(2):249–261. doi: 10.1016/0092-8674(78)90194-0. [DOI] [PubMed] [Google Scholar]
  22. Graessmann M., Graessman A. "Early" simian-virus-40-specific RNA contains information for tumor antigen formation and chromatin replication. Proc Natl Acad Sci U S A. 1976 Feb;73(2):366–370. doi: 10.1073/pnas.73.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Henderson D., Weber K. Immuno-electron microscopical identification of the two types of intermediate filaments in established epithelial cells. Exp Cell Res. 1981 Apr;132(2):297–311. doi: 10.1016/0014-4827(81)90106-3. [DOI] [PubMed] [Google Scholar]
  24. Horwitz B., Kupfer H., Eshhar Z., Geiger B. Reorganization of arrays of prekeratin filaments during mitosis. Immunofluorescence microscopy with multiclonal and monoclonal prekeratin antibodies. Exp Cell Res. 1981 Aug;134(2):281–290. doi: 10.1016/0014-4827(81)90427-4. [DOI] [PubMed] [Google Scholar]
  25. Hynes R. O., Destree A. T. 10 nm filaments in normal and transformed cells. Cell. 1978 Jan;13(1):151–163. doi: 10.1016/0092-8674(78)90146-0. [DOI] [PubMed] [Google Scholar]
  26. Jackson B. W., Grund C., Schmid E., Bürki K., Franke W. W., Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. Intermediate filaments of the cytokeratin type and desmosomes in preimplantation embryos. Differentiation. 1980;17(3):161–179. doi: 10.1111/j.1432-0436.1980.tb01093.x. [DOI] [PubMed] [Google Scholar]
  27. Jackson B. W., Grund C., Winter S., Franke W. W., Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. II. Epithelial differentiation and intermediate-sized filaments in early postimplantation embryos. Differentiation. 1981;20(3):203–216. doi: 10.1111/j.1432-0436.1981.tb01177.x. [DOI] [PubMed] [Google Scholar]
  28. Klymkowsky M. W. Intermediate filaments in 3T3 cells collapse after intracellular injection of a monoclonal anti-intermediate filament antibody. Nature. 1981 May 21;291(5812):249–251. doi: 10.1038/291249a0. [DOI] [PubMed] [Google Scholar]
  29. Klymkowsky M. W. Vimentin and keratin intermediate filament systems in cultured PtK2 epithelial cells are interrelated. EMBO J. 1982;1(2):161–165. doi: 10.1002/j.1460-2075.1982.tb01141.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lane E. B., Klymkowsky M. W. Epithelial tonofilaments: investigating their form and function using monoclonal antibodies. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):387–402. doi: 10.1101/sqb.1982.046.01.038. [DOI] [PubMed] [Google Scholar]
  31. Lane E. B. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol. 1982 Mar;92(3):665–673. doi: 10.1083/jcb.92.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  33. Lee L. D., Baden H. P. Organisation of the polypeptide chains in mammalian keratin. Nature. 1976 Nov 25;264(5584):377–379. doi: 10.1038/264377a0. [DOI] [PubMed] [Google Scholar]
  34. Lee S. Y., Krsmanovic V., Brawerman G. Attachment of ribosomes to membranes during polysome formation in mouse sarcoma 180 cells. J Cell Biol. 1971 Jun;49(3):683–691. doi: 10.1083/jcb.49.3.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lenk R., Ransom L., Kaufmann Y., Penman S. A cytoskeletal structure with associated polyribosomes obtained from HeLa cells. Cell. 1977 Jan;10(1):67–78. doi: 10.1016/0092-8674(77)90141-6. [DOI] [PubMed] [Google Scholar]
  36. Lin J. J., Feramisco J. R. Disruption of the in vivo distribution of the intermediate filaments in fibroblasts through the microinjection of a specific monoclonal antibody. Cell. 1981 Apr;24(1):185–193. doi: 10.1016/0092-8674(81)90514-6. [DOI] [PubMed] [Google Scholar]
  37. OVERTON J. Desmosome development in normal and reassociating cells in the early chick blastoderm. Dev Biol. 1962 Jun;4:532–548. doi: 10.1016/0012-1606(62)90056-8. [DOI] [PubMed] [Google Scholar]
  38. Osborn M., Franke W., Weber K. Direct demonstration of the presence of two immunologically distinct intermediate-sized filament systems in the same cell by double immunofluorescence microscopy. Vimentin and cytokeratin fibers in cultured epithelial cells. Exp Cell Res. 1980 Jan;125(1):37–46. doi: 10.1016/0014-4827(80)90186-x. [DOI] [PubMed] [Google Scholar]
  39. Paulin D., Babinet C., Weber K., Osborn M. Antibodies as probes of cellular differentiation and cytoskeletal organization in the mouse blastocyst. Exp Cell Res. 1980 Dec;130(2):297–304. doi: 10.1016/0014-4827(80)90006-3. [DOI] [PubMed] [Google Scholar]
  40. Pruss R. M., Mirsky R., Raff M. C., Thorpe R., Dowding A. J., Anderton B. H. All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell. 1981 Dec;27(3 Pt 2):419–428. doi: 10.1016/0092-8674(81)90383-4. [DOI] [PubMed] [Google Scholar]
  41. Schliwa M., Euteneuer U. Structural transformation of epidermal tonofilaments upon cold treatment. Exp Cell Res. 1979 Aug;122(1):93–101. doi: 10.1016/0014-4827(79)90564-0. [DOI] [PubMed] [Google Scholar]
  42. Sheterline P. Localisation of the major high-molecular-weight protein on microtubules in vitro and in cultured cells. Exp Cell Res. 1978 Sep;115(2):460–464. doi: 10.1016/0014-4827(78)90310-5. [DOI] [PubMed] [Google Scholar]
  43. Solomon F. Detailed neurite morphologies of sister neurolbastoma cells are related. Cell. 1979 Jan;16(1):165–169. doi: 10.1016/0092-8674(79)90197-1. [DOI] [PubMed] [Google Scholar]
  44. Stacey D. W., Allfrey V. G. Evidence for the autophagy of microinjected proteins in HeLA cells. J Cell Biol. 1977 Dec;75(3):807–817. doi: 10.1083/jcb.75.3.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Stacey D. W., Allfrey V. G. Microinjection studies of duck globin messenger RNA translation in human and avian cells. Cell. 1976 Dec;9(4 Pt 2):725–732. doi: 10.1016/0092-8674(76)90136-7. [DOI] [PubMed] [Google Scholar]
  46. Steinert P. M., Idler W. W., Goldman R. D. Intermediate filaments of baby hamster kidney (BHK-21) cells and bovine epidermal keratinocytes have similar ultrastructures and subunit domain structures. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4534–4538. doi: 10.1073/pnas.77.8.4534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Steinert P. M., Idler W. W., Zimmerman S. B. Self-assembly of bovine epidermal keratin filaments in vitro. J Mol Biol. 1976 Dec 15;108(3):547–567. doi: 10.1016/s0022-2836(76)80136-2. [DOI] [PubMed] [Google Scholar]
  48. WALEN K. H., BROWN S. W. Chromosomes in a marsupial (Potorous tridactylis) tissue culture. Nature. 1962 Apr 28;194:406–406. doi: 10.1038/194406a0. [DOI] [PubMed] [Google Scholar]
  49. Wood J. N., Anderton B. H. Monoclonal antibodies to mammalian neurofilaments. Biosci Rep. 1981 Mar;1(3):263–268. doi: 10.1007/BF01114913. [DOI] [PubMed] [Google Scholar]
  50. Zerban H., Franke W. W. Modified desmosomes in cultured epithelial cells. Cytobiologie. 1978 Dec;18(2):360–373. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES