Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Aug 1;86(2):475–482. doi: 10.1083/jcb.86.2.475

Localization of myosin, actin, and tropomyosin in rat intestinal epithelium: immunohistochemical studies at the light and electron microscope levels

PMCID: PMC2111486  PMID: 6893199

Abstract

Myosin, tropomyosin, and actin were localized in the epithelial cells of rat intestine by means of specific antibodies to chicken gizzard smooth muscle myosin, tropomyosin, and actin by immunohistochemical studies at both the light and electron microscope levels (unlabeled antibody enzyme technique). The pattern of antibody staining was the following (a) Anti-actin was associated with the microfilament bundles of the microvilli in their entire length, as well as with the microfilament network in the terminal web. (b) Anti-myosin was concentrated along the rootlets of the microvillar microfilament bundles and within the filamentous feltwork forming the terminal web. (c) Anti-tropomyosin showed a distribution similar to that of anti- myosin. In addition, the three antibodies also labeled the subplasmalemmal web underneath the cell membrane bordering on the basal lamina. Utilizing the above ultrastructural findings, we wish to propose a functional model of microvillar contraction.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. 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]
  2. Booyse F. M., Sternberger L. A., Zschocke D., Rafelson M. E., Jr Ultrastructural localization of contractile protein (thrombosthenin) in human platelets using an unlabeled antibody-peroxidase staining technique. J Histochem Cytochem. 1971 Sep;19(9):540–550. doi: 10.1177/19.9.540. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bretscher A., Weber K. Purification of microvilli and an analysis of the protein components of the microfilament core bundle. Exp Cell Res. 1978 Oct 15;116(2):397–407. doi: 10.1016/0014-4827(78)90463-9. [DOI] [PubMed] [Google Scholar]
  5. Brunser O., Luft H. J. Fine structure of the apex of absorptive cell from rat small intestine. J Ultrastruct Res. 1970 May;31(3):291–311. doi: 10.1016/s0022-5320(70)90133-4. [DOI] [PubMed] [Google Scholar]
  6. Burkl B., Mahlmeister C., Gröschel-Stewart U., Chamley-Campbell J., Campbell G. Production of specific antibodies to contractile proteins and their use in immunofluorescence microscopy. III. Antiobody against human uterine smooth muscle myosin. Histochemistry. 1979 Apr 3;60(2):135–143. doi: 10.1007/BF00495749. [DOI] [PubMed] [Google Scholar]
  7. Chamley-Campbell J., Campbell G. R., Gröschel-Stewart U., Burnstock G. FITC-labelled antibody staining of tropomyosin-containing fibrils in smooth, cardiac and skeletal muscle cells, prefusion myoblasts, fibroblasts, endothelial cells and 3T3 cells in culture. Cell Tissue Res. 1977 Sep 26;183(2):153–166. doi: 10.1007/BF00226616. [DOI] [PubMed] [Google Scholar]
  8. Chamley-Campbell J., Campbell G. R., Ross R. The smooth muscle cell in culture. Physiol Rev. 1979 Jan;59(1):1–61. doi: 10.1152/physrev.1979.59.1.1. [DOI] [PubMed] [Google Scholar]
  9. Chamley J. H., Gröschel-Stewart U., Campbell G. R., Burnstock G. Distinction between smooth muscle, fibroblasts and endothelial cells in culture by the use of fluoresceinated antibodies against smooth muscle actin. Cell Tissue Res. 1977 Feb 14;177(4):445–457. doi: 10.1007/BF00220606. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Drenckhahn D., Gröschel-Stewart U. Localization of myosin and actin in ocular nonmuscle cells. Immunofluorescence-microscopic, biochemical, and electron-microscopic studies. Cell Tissue Res. 1977 Jul 19;181(4):493–503. doi: 10.1007/BF00221771. [DOI] [PubMed] [Google Scholar]
  12. Drenckhahn D., Gröschel-Stewart U., Unsicker K. Immunofluorescence-microscopic demonstration of myosin and actin in salivary glands and exocrine pancreas of the rat. Cell Tissue Res. 1977 Sep 26;183(2):273–279. doi: 10.1007/BF00226624. [DOI] [PubMed] [Google Scholar]
  13. Drenckhahn D., Steffens R., Gröschel-Stewart U. Immunocytochemical localization of myosin in the brush border region of the intestinal epithelium. Cell Tissue Res. 1980;205(1):163–166. doi: 10.1007/BF00234452. [DOI] [PubMed] [Google Scholar]
  14. Gabbiani G., Montesano R., Tuchweber B., Salas M., Orci L. Phalloidin-induced hyperplasia of actin filaments in rat hepatocytes. Lab Invest. 1975 Nov;33(5):562–569. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Gröschel-Stewart U., Schreiber J., Mahlmeister C. Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated chicken muscle myosins. Histochemistry. 1976 Feb 26;46(3):229–236. doi: 10.1007/BF02462786. [DOI] [PubMed] [Google Scholar]
  18. Harboe N., Ingild A. Immunization, isolation of immunoglobulins, estimation of antibody titre. Scand J Immunol Suppl. 1973;1:161–164. doi: 10.1111/j.1365-3083.1973.tb03798.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Ishikawa H., Bischoff R., Holtzer H. Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol. 1969 Nov;43(2):312–328. [PMC free article] [PubMed] [Google Scholar]
  21. LEBLOND C. P., MESSIER B. Renewal of chief cells and goblet cells in the small intestine as shown by radioautography after injection of thymidine-H3 into mice. Anat Rec. 1958 Nov;132(3):247–259. doi: 10.1002/ar.1091320303. [DOI] [PubMed] [Google Scholar]
  22. Lazarides E. Two general classes of cytoplasmic actin filaments in tissue culture cells: the role of tropomyosin. J Supramol Struct. 1976;5(4):531(383)–563(415). doi: 10.1002/jss.400050410. [DOI] [PubMed] [Google Scholar]
  23. Mooseker M. S. Brush border motility. Microvillar contraction in triton-treated brush borders isolated from intestinal epithelium. J Cell Biol. 1976 Nov;71(2):417–433. doi: 10.1083/jcb.71.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Mooseker M. S., Tilney L. G. Organization of an actin filament-membrane complex. Filament polarity and membrane attachment in the microvilli of intestinal epithelial cells. J Cell Biol. 1975 Dec;67(3):725–743. doi: 10.1083/jcb.67.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pollard T. D., Korn E. D. Acanthamoeba myosin. I. Isolation from Acanthamoeba castellanii of an enzyme similar to muscle myosin. J Biol Chem. 1973 Jul 10;248(13):4682–4690. [PubMed] [Google Scholar]
  27. 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]
  28. Silverman A. J. Ultrastructural studies on the localization of neurohypophysial hormones and their carrier proteins. J Histochem Cytochem. 1976 Jul;24(7):816–827. doi: 10.1177/24.7.60434. [DOI] [PubMed] [Google Scholar]
  29. Sternberger L. A., Hardy P. H., Jr, Cuculis J. J., Meyer H. G. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem. 1970 May;18(5):315–333. doi: 10.1177/18.5.315. [DOI] [PubMed] [Google Scholar]

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

RESOURCES