Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Aug 1;103(2):613–619. doi: 10.1083/jcb.103.2.613

Myosin at the apical pole of ciliated epithelial cells as revealed by a monoclonal antibody

PMCID: PMC2113832  PMID: 3525577

Abstract

A monoclonal antibody (CC-212), obtained in a fusion experiment in which basal bodies from quail oviduct were used as immunogen, has been shown to label the apical pole of ciliated cells and to react with a 200-kD protein. This monoclonal antibody was demonstrated to be an anti- myosin from smooth muscle or from nonmuscular cells using the following criteria: On Western blots it reacted with the myosin heavy chains from gizzard and platelet extracts and from cultured cell line extracts, but did not react with striated muscle myosin heavy chains. By immunofluorescence it decorated the stress fibers of well-spread cells with a characteristic striated pattern, while it did not react with myotubes containing organized myofibrils. On native ciliated cells as well as on Triton-extracted ciliated cortices from quail oviduct, this monoclonal antibody decorated the apical pole with a stronger labeling of the periphery of the apical area. Ultrastructural localization was attempted using the immunogold technique on the same preparation. Myosin was associated with a filamentous material present between striated rootlets and the proximal extremities of the basal bodies. No labeling of the basal body itself or of axoneme was observed.

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. Anderson R. G. Biochemical and cytochemical evidence for ATPase activity in basal bodies isolated from oviduct. J Cell Biol. 1977 Aug;74(2):547–560. doi: 10.1083/jcb.74.2.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson R. G., Floyd A. K. Adenosine triphosphate induced change in light scattering of isolated basal bodies. Cell Biol Int Rep. 1978 Sep;2(5):487–494. doi: 10.1016/0309-1651(78)90101-7. [DOI] [PubMed] [Google Scholar]
  3. Biozzi G., Mouton D., Bouthillier Y., Mevel J. C. Amplification des caractères "bonne production" ou "mauvaise production" d'anticorps, produits par sélection génétique chez la souris. Corrélation entre l'immunodéficience et l'incidence de lymphomes. C R Acad Sci III. 1984;299(19):753–758. [PubMed] [Google Scholar]
  4. Bretscher A., Lynch W. Identification and localization of immunoreactive forms of caldesmon in smooth and nonmuscle cells: a comparison with the distributions of tropomyosin and alpha-actinin. J Cell Biol. 1985 May;100(5):1656–1663. doi: 10.1083/jcb.100.5.1656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Chaitin M. H., Schneider B. G., Hall M. O., Papermaster D. S. Actin in the photoreceptor connecting cilium: immunocytochemical localization to the site of outer segment disk formation. J Cell Biol. 1984 Jul;99(1 Pt 1):239–247. doi: 10.1083/jcb.99.1.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clarke M., Spudich J. A. Nonmuscle contractile proteins: the role of actin and myosin in cell motility and shape determination. Annu Rev Biochem. 1977;46:797–822. doi: 10.1146/annurev.bi.46.070177.004053. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Courvalin J. C., Dumontier M., Bornens M. Solubilization of nuclear structures by the polyanion heparin. J Biol Chem. 1982 Jan 10;257(1):456–463. [PubMed] [Google Scholar]
  10. Fujiwara K., Pollard T. D. Fluorescent antibody localization of myosin in the cytoplasm, cleavage furrow, and mitotic spindle of human cells. J Cell Biol. 1976 Dec;71(3):848–875. doi: 10.1083/jcb.71.3.848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  12. Gordon R. E., Lane B. P., Miller F. Identification of contractile proteins in basal bodies of ciliated tracheal epithelial cells. J Histochem Cytochem. 1980 Nov;28(11):1189–1197. doi: 10.1177/28.11.7000888. [DOI] [PubMed] [Google Scholar]
  13. Herman I. M., Crisona N. J., Pollard T. D. Relation between cell activity and the distribution of cytoplasmic actin and myosin. J Cell Biol. 1981 Jul;90(1):84–91. doi: 10.1083/jcb.90.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Klotz C., Bordes N., Laine M. C., Sandoz D., Bornens M. A protein of 175,000 daltons associated with striated rootlets in ciliated epithelia, as revealed by a monoclonal antibody. Cell Motil Cytoskeleton. 1986;6(1):56–67. doi: 10.1002/cm.970060108. [DOI] [PubMed] [Google Scholar]
  16. Korn E. D. Biochemistry of actomyosin-dependent cell motility (a review). Proc Natl Acad Sci U S A. 1978 Feb;75(2):588–599. doi: 10.1073/pnas.75.2.588. [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. Mooseker M. S. Actin binding proteins of the brush border. Cell. 1983 Nov;35(1):11–13. doi: 10.1016/0092-8674(83)90202-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  21. Owaribe K., Kodama R., Eguchi G. Demonstration of contractility of circumferential actin bundles and its morphogenetic significance in pigmented epithelium in vitro and in vivo. J Cell Biol. 1981 Aug;90(2):507–514. doi: 10.1083/jcb.90.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. PERRY S. V. The bound nucleotide of the isolated myofibril. Biochem J. 1952 Jul;51(4):495–499. doi: 10.1042/bj0510495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Phillips D. R., Jennings L. K., Edwards H. H. Identification of membrane proteins mediating the interaction of human platelets. J Cell Biol. 1980 Jul;86(1):77–86. doi: 10.1083/jcb.86.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  25. Porzio M. A., Pearson A. M. Improved resolution of myofibrillar proteins with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochim Biophys Acta. 1977 Jan 25;490(1):27–34. doi: 10.1016/0005-2795(77)90102-7. [DOI] [PubMed] [Google Scholar]
  26. Reed W., Avolio J., Satir P. The cytoskeleton of the apical border of the lateral cells of freshwater mussel gill: structural integration of microtubule and actin filament-based organelles. J Cell Sci. 1984 Jun;68:1–33. doi: 10.1242/jcs.68.1.1. [DOI] [PubMed] [Google Scholar]
  27. Reverdin N., Gabbiani G., Kapanci Y. Actin in tracheo-bronchial ciliated epithelial cells. Experientia. 1975 Nov 15;31(11):1348–1350. doi: 10.1007/BF01945820. [DOI] [PubMed] [Google Scholar]
  28. Sandoz D., Gounon P., Karsenti E., Sauron M. E. Immunocytochemical localization of tubulin, actin, and myosin in axonemes of ciliated cells from quail oviduct. Proc Natl Acad Sci U S A. 1982 May;79(10):3198–3202. doi: 10.1073/pnas.79.10.3198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sobieszek A., Bremel R. D. Preparation and properties of vertebrate smooth-muscle myofibrils and actomyosin. Eur J Biochem. 1975 Jun 16;55(1):49–60. doi: 10.1111/j.1432-1033.1975.tb02137.x. [DOI] [PubMed] [Google Scholar]
  30. Tassin A. M., Maro B., Bornens M. Fate of microtubule-organizing centers during myogenesis in vitro. J Cell Biol. 1985 Jan;100(1):35–46. doi: 10.1083/jcb.100.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weber K., Groeschel-Stewart U. Antibody to myosin: the specific visualization of myosin-containing filaments in nonmuscle cells. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4561–4564. doi: 10.1073/pnas.71.11.4561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weeds A. Actin-binding proteins--regulators of cell architecture and motility. Nature. 1982 Apr 29;296(5860):811–816. doi: 10.1038/296811a0. [DOI] [PubMed] [Google Scholar]
  33. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]
  34. Yerna M. J., Dabrowska R., Hartshorne D. J., Goldman R. D. Calcium-sensitive regulation of actin-myosin interactions in baby hamster kidney (BHK-21) cells. Proc Natl Acad Sci U S A. 1979 Jan;76(1):184–188. doi: 10.1073/pnas.76.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES