Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1987 Dec;129(3):589–600.

Characterization of intestinal brush border cytoskeletal proteins of normal and neoplastic human epithelial cells. A comparison with the avian brush border.

J M Carboni 1, C L Howe 1, A B West 1, K W Barwick 1, M S Mooseker 1, J S Morrow 1
PMCID: PMC1899811  PMID: 3425692

Abstract

The elaborate cytoskeletal matrix underlying the intestinal epithelial cell brush border (BB) is the hallmark of a mature enterocyte. As such, alterations in this structure are potentially useful as markers aiding in the recognition of subtle defects in cell maturation, such as those accompanying dysplasia and neoplasia. For exploration of this hypothesis, the BB components of human ileal and colonic enterocytes have been compared structurally and biochemically with the well-characterized avian BB, and alterations in the BB cytoskeleton in various states of dysplasia and neoplasia have been identified. Ultrastructural analysis of isolated human ileal BBs indicate that the human BB is structurally homologous to BBs isolated from chicken and other mammalian sources. Like other mammalian BBs (eg, from rat) the terminal web cytoskeleton of the human BB is less extensive than that in the avian BB. Immunochemical analysis of isolated human BBs indicates that the major proteins of the avian microvillar actin bundle, villin, fimbrin, and the 110-kd subunit of the 110K-calmodulin complex, are all present in the human BB. The terminal web protein myosin is also present. Unlike the terminal web of the avian BB, which contains a BB-specific isoform of spectrin, TW 260/240, the human BB contains the more widely distributed spectrin isoform, fodrin. In addition, the human BB contains multiple proteins immunoreactive with antibodies to protein 4.1, a spectrin/actin binding protein that is absent from the avian BB. Immunolocalization studies examining the distribution of the BB-specific microvillar protein, villin, in human colonic mucosa indicate that the localization of this protein is disrupted in certain dysplastic and neoplastic states. Thus, both the expression and/or distribution of BB-specific proteins such as villin may be useful markers for defects in the differentiation state of the enterocyte.

Full text

PDF
589

Images in this article

592Figure 1
on p.592
593Figure 2
on p.593
594Figure 3
on p.594
595Figure 4
on p.595
595Figure 5
on p.595
595Figure 6
on p.595
596Figure 7
on p.596
597Figure 8
on p.597
598Figure 9
on p.598

Selected References

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

  1. Bennett V. The membrane skeleton of human erythrocytes and its implications for more complex cells. Annu Rev Biochem. 1985;54:273–304. doi: 10.1146/annurev.bi.54.070185.001421. [DOI] [PubMed] [Google Scholar]
  2. Coleman T. R., Harris A. S., Mische S. M., Mooseker M. S., Morrow J. S. Beta spectrin bestows protein 4.1 sensitivity on spectrin-actin interactions. J Cell Biol. 1987 Mar;104(3):519–526. doi: 10.1083/jcb.104.3.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins J. H., Borysenko C. W. The 110,000-dalton actin- and calmodulin-binding protein from intestinal brush border is a myosin-like ATPase. J Biol Chem. 1984 Nov 25;259(22):14128–14135. [PubMed] [Google Scholar]
  4. Conzelman K. A., Mooseker M. S. The 110-kD protein-calmodulin complex of the intestinal microvillus is an actin-activated MgATPase. J Cell Biol. 1987 Jul;105(1):313–324. doi: 10.1083/jcb.105.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Croall D. E., Morrow J. S., DeMartino G. N. Limited proteolysis of the erythrocyte membrane skeleton by calcium-dependent proteinases. Biochim Biophys Acta. 1986 Jul 16;882(3):287–296. doi: 10.1016/0304-4165(86)90250-3. [DOI] [PubMed] [Google Scholar]
  6. Franke W. W., Winter S., Grund C., Schmid E., Schiller D. L., Jarasch E. D. Isolation and characterization of desmosome-associated tonofilaments from rat intestinal brush border. J Cell Biol. 1981 Jul;90(1):116–127. doi: 10.1083/jcb.90.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gerke V., Weber K. Isolation and characterization of mammalian villin and fimbrin, the two bundling proteins of the intestinal microvilli. Eur J Cell Biol. 1983 Sep;31(2):249–255. [PubMed] [Google Scholar]
  8. Glenney J. R., Jr, Glenney P. Spectrin, fodrin, and TW260/240: a family of related proteins lining the plasma membrane. Cell Motil. 1983;3(5-6):671–682. doi: 10.1002/cm.970030531. [DOI] [PubMed] [Google Scholar]
  9. Gröne H. J., Weber K., Helmchen U., Osborn M. Villin--a marker of brush border differentiation and cellular origin in human renal cell carcinoma. Am J Pathol. 1986 Aug;124(2):294–302. [PMC free article] [PubMed] [Google Scholar]
  10. Harris A. S., Anderson J. P., Yurchenco P. D., Green L. A., Ainger K. J., Morrow J. S. Mechanisms of cytoskeletal regulation: functional and antigenic diversity in human erythrocyte and brain beta spectrin. J Cell Biochem. 1986;30(1):51–69. doi: 10.1002/jcb.240300107. [DOI] [PubMed] [Google Scholar]
  11. Hirokawa N., Cheney R. E., Willard M. Location of a protein of the fodrin-spectrin-TW260/240 family in the mouse intestinal brush border. Cell. 1983 Mar;32(3):953–965. doi: 10.1016/0092-8674(83)90080-6. [DOI] [PubMed] [Google Scholar]
  12. Howe C. L., Keller T. C., 3rd, Mooseker M. S., Wasserman R. H. Analysis of cytoskeletal proteins and Ca2+-dependent regulation of structure in intestinal brush borders from rachitic chicks. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1134–1138. doi: 10.1073/pnas.79.4.1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Howe C. L., Mooseker M. S. Characterization of the 110-kdalton actin-calmodulin-, and membrane-binding protein from microvilli of intestinal epithelial cells. J Cell Biol. 1983 Oct;97(4):974–985. doi: 10.1083/jcb.97.4.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Howe C. L., Sacramone L. M., Mooseker M. S., Morrow J. S. Mechanisms of cytoskeletal regulation: modulation of membrane affinity in avian brush border and erythrocyte spectrins. J Cell Biol. 1985 Oct;101(4):1379–1385. doi: 10.1083/jcb.101.4.1379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Keller T. C., 3rd, Conzelman K. A., Chasan R., Mooseker M. S. Role of myosin in terminal web contraction in isolated intestinal epithelial brush borders. J Cell Biol. 1985 May;100(5):1647–1655. doi: 10.1083/jcb.100.5.1647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Keller T. C., 3rd, Mooseker M. S. Ca++-calmodulin-dependent phosphorylation of myosin, and its role in brush border contraction in vitro. J Cell Biol. 1982 Dec;95(3):943–959. doi: 10.1083/jcb.95.3.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Madri J. A., Barwick K. W. An immunohistochemical study of nasopharyngeal neoplasms using keratin antibodies: epithelial versus nonepithelial neoplasms. Am J Surg Pathol. 1982 Mar;6(2):143–149. doi: 10.1097/00000478-198203000-00006. [DOI] [PubMed] [Google Scholar]
  20. Matsudaira P., Mandelkow E., Renner W., Hesterberg L. K., Weber K. Role of fimbrin and villin in determining the interfilament distances of actin bundles. Nature. 1983 Jan 20;301(5897):209–214. doi: 10.1038/301209a0. [DOI] [PubMed] [Google Scholar]
  21. Mooseker M. S., Coleman T. R., Conzelman K. A. Calcium and the regulation of cytoskeletal assembly, structure and contractility. Ciba Found Symp. 1986;122:232–249. doi: 10.1002/9780470513347.ch14. [DOI] [PubMed] [Google Scholar]
  22. Mooseker M. S., Howe C. L. The brush border of intestinal epithelium: a model system for analysis of cell-surface architecture and motility. Methods Cell Biol. 1982;25(Pt B):143–174. doi: 10.1016/s0091-679x(08)61424-7. [DOI] [PubMed] [Google Scholar]
  23. Mooseker M. S. Organization, chemistry, and assembly of the cytoskeletal apparatus of the intestinal brush border. Annu Rev Cell Biol. 1985;1:209–241. doi: 10.1146/annurev.cb.01.110185.001233. [DOI] [PubMed] [Google Scholar]
  24. Pearl M., Fishkind D., Mooseker M., Keene D., Keller T., 3rd Studies on the spectrin-like protein from the intestinal brush border, TW 260/240, and characterization of its interaction with the cytoskeleton and actin. J Cell Biol. 1984 Jan;98(1):66–78. doi: 10.1083/jcb.98.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robine S., Huet C., Moll R., Sahuquillo-Merino C., Coudrier E., Zweibaum A., Louvard D. Can villin be used to identify malignant and undifferentiated normal digestive epithelial cells? Proc Natl Acad Sci U S A. 1985 Dec;82(24):8488–8492. doi: 10.1073/pnas.82.24.8488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rodewald R., Newman S. B., Karnovsky M. J. Contraction of isolated brush borders from the intestinal epithelium. J Cell Biol. 1976 Sep;70(3):541–554. doi: 10.1083/jcb.70.3.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rodman J. S., Mooseker M., Farquhar M. G. Cytoskeletal proteins of the rat kidney proximal tubule brush border. Eur J Cell Biol. 1986 Dec;42(2):319–327. [PubMed] [Google Scholar]
  28. Semenza G. Anchoring and biosynthesis of stalked brush border membrane proteins: glycosidases and peptidases of enterocytes and renal tubuli. Annu Rev Cell Biol. 1986;2:255–313. doi: 10.1146/annurev.cb.02.110186.001351. [DOI] [PubMed] [Google Scholar]
  29. Shibayama T., Carboni J. M., Mooseker M. S. Assembly of the intestinal brush border: appearance and redistribution of microvillar core proteins in developing chick enterocytes. J Cell Biol. 1987 Jul;105(1):335–344. doi: 10.1083/jcb.105.1.335. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES