Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Aug 1;86(2):568–575. doi: 10.1083/jcb.86.2.568

Banding and polarity of actin filaments in interphase and cleaving cells

PMCID: PMC2111493  PMID: 6995468

Abstract

Heavy meromyosin (HMM) decoration of actin filaments was used to detect the polarity of microfilaments in interphase and cleaving rat kangaroo (PtK2) cells. Ethanol at -20 degrees C was used to make the cells permeable to HMM followed by tannic acid-glutaraldehyde fixation for electron microscopy. Uniform polarity of actin filaments was observed at cell junctions and central attachment plaques with the HMM arrowheads always pointing away from the junction or plaque. Stress fibers were banded in appearance with their component microfilaments exhibiting both parallel and antiparallel orientation with respect to one another. Identical banding of microfilament bundles was also seen in cleavage furrows with the same variation in filament polarity as found in stress fibers. Similarly banded fibers were not seen outside the cleavage furrow in mitotic cells. By the time that a mid-body was present, the actin filaments in the cleavage furrow were no longer in banded fibers. The alternating dark and light bands of both the stress fibers and cleavage furrow fibers are approximately equal in length, each measuring approximately 0.16 micrometer. Actin filaments were present in both bands, and individual decorated filaments could sometimes be traced through four band lengths. Undecorated filaments, 10 nm in diameter, could often be seen within the light bands. A model is proposed to explain the arrangement of filaments in stress fibers and cleavage furrows based on the striations observed with tannic acid and the polarity of the actin filaments.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. ARONSON J. OVERLAP OF THE BIREFRINGENT COMPONENT OF ADJACENT A REGIONS DURING THE INDUCED SHORTENING OF FIBRILS TEASED FROM DROSOPHILA MUSCLE. J Cell Biol. 1963 Oct;19:107–114. doi: 10.1083/jcb.19.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. IV. Electron microscopy of the leading lamella. Exp Cell Res. 1971 Aug;67(2):359–367. doi: 10.1016/0014-4827(71)90420-4. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Buckley I. K., Porter K. R. Cytoplasmic fibrils in living cultured cells. A light and electron microscope study. Protoplasma. 1967;64(4):349–380. doi: 10.1007/BF01666538. [DOI] [PubMed] [Google Scholar]
  5. Burgess D. R., Schroeder T. E. Polarized bundles of actin filaments within microvilli of fertilized sea urchin eggs. J Cell Biol. 1977 Sep;74(3):1032–1037. doi: 10.1083/jcb.74.3.1032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chang C. M., Goldman R. D. The localization of actin-like fibers in cultured neuroblastoma cells as revealed by heavy meromyosin binding. J Cell Biol. 1973 Jun;57(3):867–874. doi: 10.1083/jcb.57.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edds K. T. Microfilament bundles. I. Formation with uniform polarity. Exp Cell Res. 1977 Sep;108(2):452–456. doi: 10.1016/s0014-4827(77)80056-6. [DOI] [PubMed] [Google Scholar]
  8. Forer A., Behnke O. An actin-like component in spermatocytes of a crane fly (Nephrotoma suturalis Loew). I. The spindle. Chromosoma. 1972;39(2):145–173. doi: 10.1007/BF00319840. [DOI] [PubMed] [Google Scholar]
  9. Franke W. W., Grund C., Osborn M., Weber K. The intermediate-sized filaments in rat kangaroo PtK2 cells. I. Morphology in situ. Cytobiologie. 1978 Aug;17(2):365–391. [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. Fujiwara K., Porter M. E., Pollard T. D. Alpha-actinin localization in the cleavage furrow during cytokinesis. J Cell Biol. 1978 Oct;79(1):268–275. doi: 10.1083/jcb.79.1.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gabbiani G., Elemer G., Guelpa C., Vallotton M. B., Badonnel M. C., Hüttner I. Morphologic and functional changes of the aortic intima during experimental hypertension. Am J Pathol. 1979 Aug;96(2):399–422. [PMC free article] [PubMed] [Google Scholar]
  13. Goldman R. D., Chojnacki B., Yerna M. J. Ultrastructure of microfilament bundles in baby hamster kidney (BHK-21) cells. The use of tannic acid. J Cell Biol. 1979 Mar;80(3):759–766. doi: 10.1083/jcb.80.3.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gordon W. E., 3rd Immunofluorescent and ultrastructural studies of "sarcomeric" units in stress fibers of cultured non-muscle cells. Exp Cell Res. 1978 Dec;117(2):253–260. doi: 10.1016/0014-4827(78)90138-6. [DOI] [PubMed] [Google Scholar]
  15. HUXLEY H. E. The contractile structure of cardiac and skeletal muscle. Circulation. 1961 Aug;24:328–335. doi: 10.1161/01.cir.24.2.328. [DOI] [PubMed] [Google Scholar]
  16. Hoyle G., McAlear J. H., Selverston A. Mechanism of supercontraction in a striated muscle. J Cell Biol. 1965 Aug;26(2):621–640. doi: 10.1083/jcb.26.2.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Isenberg G., Rathke P. C., Hülsmann N., Franke W. W., Wohlfarth-Bottermann K. E. Cytoplasmic actomyosin fibrils in tissue culture cells: direct proof of contractility by visualization of ATP-induced contraction in fibrils isolated by laser micro-beam dissection. Cell Tissue Res. 1976 Feb 27;166(4):427–443. doi: 10.1007/BF00225909. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. James D. W., Taylor J. F. The stress developed by sheets of chick fibroblasts in vitro. Exp Cell Res. 1969 Jan;54(1):107–110. doi: 10.1016/0014-4827(69)90299-7. [DOI] [PubMed] [Google Scholar]
  20. Lazarides E. Actin, alpha-actinin, and tropomyosin interaction in the structural organization of actin filaments in nonmuscle cells. J Cell Biol. 1976 Feb;68(2):202–219. doi: 10.1083/jcb.68.2.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lazarides E., Burridge K. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell. 1975 Nov;6(3):289–298. doi: 10.1016/0092-8674(75)90180-4. [DOI] [PubMed] [Google Scholar]
  22. Lazarides E. Tropomyosin antibody: the specific localization of tropomyosin in nonmuscle cells. J Cell Biol. 1975 Jun;65(3):549–561. doi: 10.1083/jcb.65.3.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lazarides E., Weber K. Actin antibody: the specific visualization of actin filaments in non-muscle cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2268–2272. doi: 10.1073/pnas.71.6.2268. [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. Niederman R., Pollard T. D. Human platelet myosin. II. In vitro assembly and structure of myosin filaments. J Cell Biol. 1975 Oct;67(1):72–92. doi: 10.1083/jcb.67.1.72. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pepe F. A. Structure of muscle filaments from immunohistochemical and ultrastructural studies. J Histochem Cytochem. 1975 Jul;23(7):543–562. doi: 10.1177/23.7.1095653. [DOI] [PubMed] [Google Scholar]
  28. Robinson T. F., Winegrad S. The measurement and dynamic implications of thin filament lengths in heart muscle. J Physiol. 1979 Jan;286:607–619. doi: 10.1113/jphysiol.1979.sp012640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Röhlich P., Oláh I. Cross-striated fibrils in the endothelium of the rat myometral arterioles. J Ultrastruct Res. 1967 Jun;18(5):667–676. doi: 10.1016/s0022-5320(67)80212-0. [DOI] [PubMed] [Google Scholar]
  30. Sanger J. W. Intracellular localization of actin with fluorescently labelled heavy meromyosin. Cell Tissue Res. 1975 Aug 27;161(4):431–434. doi: 10.1007/BF00224134. [DOI] [PubMed] [Google Scholar]
  31. Sanger J. W. Presence of actin during chromosomal movement. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2451–2455. doi: 10.1073/pnas.72.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanger J. W., Sanger J. M. The cytoskeleton and cell division. Methods Achiev Exp Pathol. 1979;8:110–142. [PubMed] [Google Scholar]
  33. Schroeder T. E. Cytokinesis: filaments in the cleavage furrow. Exp Cell Res. 1968 Oct;53(1):272–276. doi: 10.1016/0014-4827(68)90373-x. [DOI] [PubMed] [Google Scholar]
  34. Seagull R. W., Heath I. B. The effects of tannic acid on the in vivo preservation of microfilaments. Eur J Cell Biol. 1979 Dec;20(2):184–188. [PubMed] [Google Scholar]
  35. 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]
  36. Small J. V., Isenberg G., Celis J. E. Polarity of actin at the leading edge of cultured cells. Nature. 1978 Apr 13;272(5654):638–639. doi: 10.1038/272638a0. [DOI] [PubMed] [Google Scholar]
  37. Szent-Györgyi A. G., Cohen C., Kendrick-Jones J. Paramyosin and the filaments of molluscan "catch" muscles. II. Native filaments: isolation and characterization. J Mol Biol. 1971 Mar 14;56(2):239–258. doi: 10.1016/0022-2836(71)90462-1. [DOI] [PubMed] [Google Scholar]
  38. Tilney L. G., Kallenbach N. Polymerization of actin. VI. The polarity of the actin filaments in the acrosomal process and how it might be determined. J Cell Biol. 1979 Jun;81(3):608–623. doi: 10.1083/jcb.81.3.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yerna M. J., Aksoy M. O., Hartshorne D. J., Goldman R. D. BHK21 myosin: isolation, biochemical characterization and intracellular localization. J Cell Sci. 1978 Jun;31:411–429. doi: 10.1242/jcs.31.1.411. [DOI] [PubMed] [Google Scholar]
  40. Zigmond S. H., Otto J. J., Bryan J. Organization of myosin in a submembranous sheath in well-spread human fibroblasts. Exp Cell Res. 1979 Mar 15;119(2):205–219. doi: 10.1016/0014-4827(79)90349-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES