Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 May 1;102(5):1843–1852. doi: 10.1083/jcb.102.5.1843

Identification of actin-, alpha-actinin-, and vinculin-containing plaques at the lateral membrane of epithelial cells

PMCID: PMC2114231  PMID: 3084501

Abstract

In this paper, a new type of spot desmosome-like junction (type II plaque) is described that is scattered along the entire lateral plasma membrane of rat and human intestinal epithelium. Ultrastructurally type II plaques differed from the classical type of epithelial spot desmosome ("macula adherens", further denoted as type I desmosome) by weak electron density of the membrane-associated plaque material, association of the plaques with microfilaments rather than intermediate filaments, and poorly visible material across the intercellular space. Thus, type II plaques resemble cross-sections of the zonula adherens. Immunofluorescence-microscopic studies were done using antibodies to a main protein associated with the plaques of type I desmosomes (desmoplakin I) and to the three major proteins located at the plaques of the zonula adherens (actin, alpha-actinin, and vinculin). Two types of plaques were visualized along the lateral surface of intestinal and prostatic epithelium: (a) the type I desmosomes, which were labeled with anti-desmoplakin but did not bind antibodies to actin, alpha- actinin, and vinculin, and (b) a further set of similarly sized plaques, which bound antibodies to actin, alpha-actinin, and vinculin but were not stained with anti-desmoplakin. Three-dimensional computer reconstruction of serial sections double-labeled with anti-desmoplakin and anti-alpha-actinin further confirmed that both types of plaques are spatially completely separated from each other along the lateral plasma membrane. The computer graphs further revealed that the actin-, alpha- actinin-, and vinculin-containing plaques have the tendency to form clusters, a feature also typical of type II plaques. It is suggested that the type II plaques represent spot desmosome-like intercellular junctions, which, like the zonula adherens, appear to be linked to the actin filament system. As the type II plaques cover a considerable part of the lateral cell surface, they might play a particular role in controlling cellular shape and intercellular adhesion.

Full Text

The Full Text of this article is available as a PDF (4.5 MB).

Selected References

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

  1. Boller K., Vestweber D., Kemler R. Cell-adhesion molecule uvomorulin is localized in the intermediate junctions of adult intestinal epithelial cells. J Cell Biol. 1985 Jan;100(1):327–332. doi: 10.1083/jcb.100.1.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bretscher A., Weber K. Localization of actin and microfilament-associated proteins in the microvilli and terminal web of the intestinal brush border by immunofluorescence microscopy. J Cell Biol. 1978 Dec;79(3):839–845. doi: 10.1083/jcb.79.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burgess D. R. Reactivation of intestinal epithelial cell brush border motility: ATP-dependent contraction via a terminal web contractile ring. J Cell Biol. 1982 Dec;95(3):853–863. doi: 10.1083/jcb.95.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Cowin P., Mattey D., Garrod D. Distribution of desmosomal components in the tissues of vertebrates, studied by fluorescent antibody staining. J Cell Sci. 1984 Mar;66:119–132. doi: 10.1242/jcs.66.1.119. [DOI] [PubMed] [Google Scholar]
  6. Craig S. W., Pardo J. V. alpha-Actinin localization in the junctional complex of intestinal epithelial cells. J Cell Biol. 1979 Jan;80(1):203–210. doi: 10.1083/jcb.80.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drenckhahn D., Frotscher M., Kaiser H. W. Concentration of F-actin in synaptic formations of the hippocampus as visualized by staining with fluorescent phalloidin. Brain Res. 1984 May 23;300(2):381–384. doi: 10.1016/0006-8993(84)90851-5. [DOI] [PubMed] [Google Scholar]
  8. Drenckhahn D., Gröschel-Stewart U. Localization of myosin, actin, and tropomyosin in rat intestinal epithelium: immunohistochemical studies at the light and electron microscope levels. J Cell Biol. 1980 Aug;86(2):475–482. doi: 10.1083/jcb.86.2.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Drenckhahn D., Hofmann H. D., Mannherz H. G. Evidence for the association of villin with core filaments and rootlets of intestinal epithelial microvilli. Cell Tissue Res. 1983;228(2):409–414. doi: 10.1007/BF00204889. [DOI] [PubMed] [Google Scholar]
  10. Drenckhahn D., Mannherz H. G. Distribution of actin and the actin-associated proteins myosin, tropomyosin, alpha-actinin, vinculin, and villin in rat and bovine exocrine glands. Eur J Cell Biol. 1983 May;30(2):167–176. [PubMed] [Google Scholar]
  11. 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]
  12. Faulstich H., Trischmann H., Mayer D. Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis. Exp Cell Res. 1983 Mar;144(1):73–82. doi: 10.1016/0014-4827(83)90443-3. [DOI] [PubMed] [Google Scholar]
  13. Feramisco J. R., Burridge K. A rapid purification of alpha-actinin, filamin, and a 130,000-dalton protein from smooth muscle. J Biol Chem. 1980 Feb 10;255(3):1194–1199. [PubMed] [Google Scholar]
  14. Franke W. W., Moll R., Mueller H., Schmid E., Kuhn C., Krepler R., Artlieb U., Denk H. Immunocytochemical identification of epithelium-derived human tumors with antibodies to desmosomal plaque proteins. Proc Natl Acad Sci U S A. 1983 Jan;80(2):543–547. doi: 10.1073/pnas.80.2.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Geiger B., Dutton A. H., Tokuyasu K. T., Singer S. J. Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells. J Cell Biol. 1981 Dec;91(3 Pt 1):614–628. doi: 10.1083/jcb.91.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Geiger B. Membrane-cytoskeleton interaction. Biochim Biophys Acta. 1983 Aug 11;737(3-4):305–341. doi: 10.1016/0304-4157(83)90005-9. [DOI] [PubMed] [Google Scholar]
  18. Geiger B., Tokuyasu K. T., Singer S. J. Immunocytochemical localization of alpha-actinin in intestinal epithelial cells. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2833–2837. doi: 10.1073/pnas.76.6.2833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorbsky G., Steinberg M. S. Isolation of the intercellular glycoproteins of desmosomes. J Cell Biol. 1981 Jul;90(1):243–248. doi: 10.1083/jcb.90.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gröschel-Stewart U., Ceurremans S., Lehr I., Mahlmeister C., Paar E. Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin. Histochemistry. 1977 Feb 1;50(4):271–279. doi: 10.1007/BF00507120. [DOI] [PubMed] [Google Scholar]
  21. Hirokawa N., Heuser J. E. Quick-freeze, deep-etch visualization of the cytoskeleton beneath surface differentiations of intestinal epithelial cells. J Cell Biol. 1981 Nov;91(2 Pt 1):399–409. doi: 10.1083/jcb.91.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hirokawa N., Keller T. C., 3rd, Chasan R., Mooseker M. S. Mechanism of brush border contractility studied by the quick-freeze, deep-etch method. J Cell Biol. 1983 May;96(5):1325–1336. doi: 10.1083/jcb.96.5.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hull B. E., Staehelin L. A. The terminal web. A reevaluation of its structure and function. J Cell Biol. 1979 Apr;81(1):67–82. doi: 10.1083/jcb.81.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. MAYOR H. D., HAMPTON J. C., ROSARIO B. A simple method for removing the resin from epoxy-embedded tissue. J Biophys Biochem Cytol. 1961 Apr;9:909–910. doi: 10.1083/jcb.9.4.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mangeat P., Burridge K. Actin-membrane interaction in fibroblasts: what proteins are involved in this association? J Cell Biol. 1984 Jul;99(1 Pt 2):95s–103s. doi: 10.1083/jcb.99.1.95s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maupin P., Pollard T. D. Improved preservation and staining of HeLa cell actin filaments, clathrin-coated membranes, and other cytoplasmic structures by tannic acid-glutaraldehyde-saponin fixation. J Cell Biol. 1983 Jan;96(1):51–62. doi: 10.1083/jcb.96.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McNutt N. S., Weinstein R. S. Membrane ultrastructure at mammalian intercellular junctions. Prog Biophys Mol Biol. 1973;26:45–101. doi: 10.1016/0079-6107(73)90017-5. [DOI] [PubMed] [Google Scholar]
  29. Merk F. B., Leav I., Kwan P. W., Ofner P. Effects of estrogen and androgen on the ultrastructure of secretory granules and intercellular junctions in regressed canine prostate. Anat Rec. 1980 Jun;197(2):111–132. doi: 10.1002/ar.1091970202. [DOI] [PubMed] [Google Scholar]
  30. Mooseker M. S., Bonder E. M., Conzelman K. A., Fishkind D. J., Howe C. L., Keller T. C., 3rd Brush border cytoskeleton and integration of cellular functions. J Cell Biol. 1984 Jul;99(1 Pt 2):104s–112s. doi: 10.1083/jcb.99.1.104s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mooseker M. S., Pollard T. D., Fujiwara K. Characterization and localization of myosin in the brush border of intestinal epithelial cells. J Cell Biol. 1978 Nov;79(2 Pt 1):444–453. doi: 10.1083/jcb.79.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. PEASE D. C., MOLINARI S. Electron microscopy of muscular arteries; pial vessels of43 the cat and monkey. J Ultrastruct Res. 1960 Jun;3:447–468. doi: 10.1016/s0022-5320(60)90022-8. [DOI] [PubMed] [Google Scholar]
  33. Pardo J. V., Siliciano J. D., Craig S. W. A vinculin-containing cortical lattice in skeletal muscle: transverse lattice elements ("costameres") mark sites of attachment between myofibrils and sarcolemma. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1008–1012. doi: 10.1073/pnas.80.4.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Russell L. Desmosome-like junctions between Sertoli and germ cells in the rat testis. Am J Anat. 1977 Mar;148(3):301–312. doi: 10.1002/aja.1001480302. [DOI] [PubMed] [Google Scholar]
  36. Skerrow C. J., Matoltsy A. G. Isolation of epidermal desmosomes. J Cell Biol. 1974 Nov;63(2 Pt 1):515–523. doi: 10.1083/jcb.63.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Staehelin L. A. Structure and function of intercellular junctions. Int Rev Cytol. 1974;39:191–283. doi: 10.1016/s0074-7696(08)60940-7. [DOI] [PubMed] [Google Scholar]
  38. Talian J. C., Olmsted J. B., Goldman R. D. A rapid procedure for preparing fluorescein-labeled specific antibodies from whole antiserum: its use in analyzing cytoskeletal architecture. J Cell Biol. 1983 Oct;97(4):1277–1282. doi: 10.1083/jcb.97.4.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Toyama Y. Actin-like filaments in the Sertoli cell junctional specializations in the swine and mouse testis. Anat Rec. 1976 Dec;186(4):477–491. doi: 10.1002/ar.1091860403. [DOI] [PubMed] [Google Scholar]
  40. Tramu G., Pillez A., Leonardelli J. An efficient method of antibody elution for the successive or simultaneous localization of two antigens by immunocytochemistry. J Histochem Cytochem. 1978 Apr;26(4):322–324. doi: 10.1177/26.4.207771. [DOI] [PubMed] [Google Scholar]

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

RESOURCES