Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Oct 1;91(1):45–54. doi: 10.1083/jcb.91.1.45

Response of basal epithelial cell surface and Cytoskeleton to solubilized extracellular matrix molecules

PMCID: PMC2111939  PMID: 7197682

Abstract

Corneal epithelium removed from underlying extracellular matrix (ECM) extends numerous cytoplasmic processes (blebs) from the formerly smooth basal surface. If blebbing epithelia are grown on collagen gels or lens capsules in vitro, the basal surface flattens and takes on the smooth contour typical of epithelium in contact with basal lamina in situ. This study examines the effect of soluble extracellular matrix components on the basal surface. Corneal epithelia from 9- to 11-d-old chick embryos were isolated with trypsin-collagenase or ethylenediamine tetraacetic acid, then placed on Millipore filters (Millipore Corp., Bedford, Mass.), and cultured at the medium-air interface. Media were prepared with no serum, with 10% of calf serum, or with serum from which plasma fibronectin was removed. Epithelia grown on filters in this medium continue to bleb for the duration of the experiments (12-14 h). If soluble collagen, laminin, or fibronectin is added to the medium, however, blebs are withdrawn and by 2-6 h the basal surface is flat. Epithelia grown on filters in the presence of albumin, IgG, or glycosaminoglycans continue to bleb. Epithelia cultured on solid substrata, such as glass, also continue to bleb if ECM is absent from the medium. The basal cell cortex in situ contains a compact cortical mat of filaments that decorate with S-1 myosin subfragments; some, if not all, of these filaments point away from the plasmalemma. The actin filaments disperse into the cytoplasmic processes during blebbing and now many appear to point toward the plasmalemma. In isolated epithelia that flatten in response to soluble collagens, laminin, and fibronectin, the actin filaments reform the basal cortical mat typical or epithelial in situ. Thus, extracellular macromolecules influence and organize not only the basal cell surface but also the actin-rich basal cell cortex of epithelial cells.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Banerjee S. D., Cohn R. H., Bernfield M. R. Basal lamina of embryonic salivary epithelia. Production by the epithelium and role in maintaining lobular morphology. J Cell Biol. 1977 May;73(2):445–463. doi: 10.1083/jcb.73.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Begg D. A., Rodewald R., Rebhun L. I. The visualization of actin filament polarity in thin sections. Evidence for the uniform polarity of membrane-associated filaments. J Cell Biol. 1978 Dec;79(3):846–852. doi: 10.1083/jcb.79.3.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berman M. D., Waggoner J. G., Foidart J. M., Kleinman H. K. Attachment to collagen by isolated hepatocytes from rats with induced hepatic fibrosis. J Lab Clin Med. 1980 May;95(5):660–671. [PubMed] [Google Scholar]
  4. Dodson J. W., Hay E. D. Secretion of collagen by corneal epithelium. II. Effect of the underlying substratum on secretion and polymerization of epithelial products. J Exp Zool. 1974 Jul;189(1):51–72. doi: 10.1002/jez.1401890106. [DOI] [PubMed] [Google Scholar]
  5. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goldberg B. Binding of soluble type I collagen molecules to the fibroblast plasma membrane. Cell. 1979 Feb;16(2):265–275. doi: 10.1016/0092-8674(79)90004-7. [DOI] [PubMed] [Google Scholar]
  7. Hasty D. L., Hay E. D. Freeze-fracture studies of the developing cell surface. II. Particle-free membrane blisters on glutaraldehyde-fixed corneal fibroblasts are artefacts. J Cell Biol. 1978 Sep;78(3):756–768. doi: 10.1083/jcb.78.3.756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hay E. D., Dodson J. W. Secretion of collagen by corneal epithelium. I. Morphology of the collagenous products produced by isolated epithelia grown on frozen-killed lens. J Cell Biol. 1973 Apr;57(1):190–213. doi: 10.1083/jcb.57.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hay E. D., Linsenmayer T. F., Trelstad R. L., von der Mark K. Origin and distribution of collagens in the developing avian cornea. Curr Top Eye Res. 1979;1:1–35. [PubMed] [Google Scholar]
  10. Hay E. D., Meier S. Glycosaminoglycan synthesis by embryonic inductors: neural tube, notochord, and lens. J Cell Biol. 1974 Sep;62(3):889–898. doi: 10.1083/jcb.62.3.889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hay E. D., Revel J. P. Fine structure of the developing avian cornea. Monogr Dev Biol. 1969;1:1–144. [PubMed] [Google Scholar]
  12. Hynes R. O., Destree A. T., Perkins M. E., Wagner D. D. Cell surface fibronectin and oncogenic transformation. J Supramol Struct. 1979;11(1):95–104. doi: 10.1002/jss.400110110. [DOI] [PubMed] [Google Scholar]
  13. Jensen H. M., Mottet N. K. Ultrastructural changes in keratinizing epithelium following trypsinization, epidermal detachment and apposition to mesenchymes. J Cell Sci. 1970 Mar;6(2):511–535. doi: 10.1242/jcs.6.2.511. [DOI] [PubMed] [Google Scholar]
  14. Kleinman H. K., Klebe R. J., Martin G. R. Role of collagenous matrices in the adhesion and growth of cells. J Cell Biol. 1981 Mar;88(3):473–485. doi: 10.1083/jcb.88.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kleinman H. K., Martin G. R., Fishman P. H. Ganglioside inhibition of fibronectin-mediated cell adhesion to collagen. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3367–3371. doi: 10.1073/pnas.76.7.3367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Linsenmayer T. F., Hendrix M. J. Monoclonal antibodies to connective tissue macromolecules: type II collagen. Biochem Biophys Res Commun. 1980 Jan 29;92(2):440–446. doi: 10.1016/0006-291x(80)90352-6. [DOI] [PubMed] [Google Scholar]
  17. Linsenmayer T. F., Smith G. N., Jr, Hay E. D. Synthesis of two collagen types by embryonic chick corneal epithelium in vitro. Proc Natl Acad Sci U S A. 1977 Jan;74(1):39–43. doi: 10.1073/pnas.74.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liotta L. A., Abe S., Robey P. G., Martin G. R. Preferential digestion of basement membrane collagen by an enzyme derived from a metastatic murine tumor. Proc Natl Acad Sci U S A. 1979 May;76(5):2268–2272. doi: 10.1073/pnas.76.5.2268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mauchamp J., Margotat A., Chambard M., Charrier B., Remy L., Michel-Bechet M. Polarity of three-dimensional structures derived from isolated hog thyroid cells in primary culture. Cell Tissue Res. 1979;204(3):417–430. doi: 10.1007/BF00233653. [DOI] [PubMed] [Google Scholar]
  20. Meier S., Hay E. D. Control of corneal differentiation by extracellular materials. Collagen as a promoter and stabilizer of epithelial stroma production. Dev Biol. 1974 Jun;38(2):249–270. doi: 10.1016/0012-1606(74)90005-0. [DOI] [PubMed] [Google Scholar]
  21. Meier S., Hay E. D. Stimulation of extracellular matrix synthesis in the developing cornea by glycosaminoglycans. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2310–2313. doi: 10.1073/pnas.71.6.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murray J. C., Stingl G., Kleinman H. K., Martin G. R., Katz S. I. Epidermal cells adhere preferentially to type IV (basement membrane) collagen. J Cell Biol. 1979 Jan;80(1):197–202. doi: 10.1083/jcb.80.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rennard S. I., Wind M. L., Hewitt A. T., Kleinman H. K. Effect of collagen and cell shape on binding of fibronectin to cells. Arch Biochem Biophys. 1981 Jan;206(1):205–212. doi: 10.1016/0003-9861(81)90082-5. [DOI] [PubMed] [Google Scholar]
  24. Rubin K., Hök M., Obrink B., Timpl R. Substrate adhesion of rat hepatocytes: mechanism of attachment to collagen substrates. Cell. 1981 May;24(2):463–470. doi: 10.1016/0092-8674(81)90337-8. [DOI] [PubMed] [Google Scholar]
  25. Rubin K., Johansson S., Pettersson I., Ocklind C., Obrink B., Hök M. Attachment of rat hepatocytes to collagen and fibronectin; a study using antibodies directed against cell surface components. Biochem Biophys Res Commun. 1979 Nov 14;91(1):86–94. doi: 10.1016/0006-291x(79)90586-2. [DOI] [PubMed] [Google Scholar]
  26. Rubin K., Oldberg A., Hök M., Obrink B. Adhesion of rat hepatocytes to collagen. Exp Cell Res. 1978 Nov;117(1):165–177. doi: 10.1016/0014-4827(78)90439-1. [DOI] [PubMed] [Google Scholar]
  27. Singer I. I. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell. 1979 Mar;16(3):675–685. doi: 10.1016/0092-8674(79)90040-0. [DOI] [PubMed] [Google Scholar]
  28. Terranova V. P., Rohrbach D. H., Martin G. R. Role of laminin in the attachment of PAM 212 (epithelial) cells to basement membrane collagen. Cell. 1980 Dec;22(3):719–726. doi: 10.1016/0092-8674(80)90548-6. [DOI] [PubMed] [Google Scholar]
  29. Timpl R., Martin G. R., Bruckner P., Wick G., Wiedemann H. Nature of the collagenous protein in a tumor basement membrane. Eur J Biochem. 1978 Mar;84(1):43–52. doi: 10.1111/j.1432-1033.1978.tb12139.x. [DOI] [PubMed] [Google Scholar]
  30. Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
  31. Trelstad R. L., Hayashi K., Toole B. P. Epithelial collagens and glycosaminoglycans in the embryonic cornea. Macromolecular order and morphogenesis in the basement membrane. J Cell Biol. 1974 Sep;62(3):815–830. doi: 10.1083/jcb.62.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Trinkaus J. P. Formation of protrusions of the cell surface during tissue cell movement. Prog Clin Biol Res. 1980;41:887–906. [PubMed] [Google Scholar]
  33. Wicha M. S., Liotta L. A., Garbisa S., Kidwell W. R. Basement membrane collagen requirements for attachment and growth of mammary epithelium. Exp Cell Res. 1979 Nov;124(1):181–190. doi: 10.1016/0014-4827(79)90268-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES