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. 1977 Mar 1;72(3):552–567. doi: 10.1083/jcb.72.3.552

A comparison of the distribution of actin and tubulin in the mammalian mitotic spindle as seen by indirect immunofluorescence

PMCID: PMC2111019  PMID: 320217

Abstract

Rabbit antibodies against actin and tubulin were used in an indirect immunofluorescence study of the structure of the mitotic spindle of PtK1 cells after lysis under conditions that preserve anaphase chromosome movement. During early prophase there is no antiactin staining associated with the mitotic centers, but by late prophase, as the spindle is beginning to form, a small ball of actin antigenicity is found beside the nucleus; After nuclear envelope breakdown, the actiactin stains the region around each mitotic center, and becomes organized into fibers that run between the chromosomes and the poles. Colchicine blocks this organization, but does not disrupt the staining at the poles. At metaphase the antiactin reveals a halo of ill-defined radius around each spindle pole and fibers that run from the poles to the metaphase plate. Antitubulin shows astral rays, fibers running from chromosomes to poles, and some fibers that run across the metaphase plate. At anaphase, there is a shortening of the antiactin-stained fibers, leaving a zone which is essentially free of actin-staining fluorescence between the separating chromosomes. Antitubulin stains the region between chromosomes and poles, but also reveals substantial fibers running through the zone between separating chromosomes. Cells fixed during cytokinesis show actin in the region of the cleavage furrow, while antitubulin reveals the fibrous spindle remnant that runs between daughter cells. These results suggest that actin is a component of the mammalian mitotic spindle, that the distribution of actin differs from that of tubulin and that the distributions of these two fibrous proteins change in different ways during anaphase.

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Selected References

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

  1. Bajer A. S. Interaction of microtubules and the mechanism of chromosome movement (zipper hypothesis). 1. General principle. Cytobios. 1973 Nov;8(31):139–160. [PubMed] [Google Scholar]
  2. Behnke O., Forer A., Emmersen J. Actin in sperm tails and meiotic spindles. Nature. 1971 Dec 17;234(5329):408–410. doi: 10.1038/234408a0. [DOI] [PubMed] [Google Scholar]
  3. Borisy G. G., Olmsted J. B. Nucleated assembly of microtubules in porcine brain extracts. Science. 1972 Sep 29;177(4055):1196–1197. doi: 10.1126/science.177.4055.1196. [DOI] [PubMed] [Google Scholar]
  4. Brinkley B. R., Cartwright J., Jr Cold-labile and cold-stable microtubules in the mitotic spindle of mammalian cells. Ann N Y Acad Sci. 1975 Jun 30;253:428–439. doi: 10.1111/j.1749-6632.1975.tb19218.x. [DOI] [PubMed] [Google Scholar]
  5. Brinkley B. R., Cartwright J., Jr Ultrastructural analysis of mitotic spindle elongation in mammalian cells in vitro. Direct microtubule counts. J Cell Biol. 1971 Aug;50(2):416–431. doi: 10.1083/jcb.50.2.416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cande W. Z., Snyder J., Smith D., Summers K., McIntosh J. R. A functional mitotic spindle prepared from mammalian cells in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1559–1563. doi: 10.1073/pnas.71.4.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cooke R., Morales M. F. Interaction of globular actin with myosin subfragments. J Mol Biol. 1971 Sep 14;60(2):249–261. doi: 10.1016/0022-2836(71)90291-9. [DOI] [PubMed] [Google Scholar]
  8. Dietz R. Die Assembly-Hypothese der Chromosomenbewegung und die Veränderungen der Spindellänge während der Anaphase I in Spermatocyten von Pales ferruginea (Tipulidae, Diptera. Chromosoma. 1972;38(1):11–76. doi: 10.1007/BF00319955. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Gawadi N. Actin in the mitotic spindle. Nature. 1971 Dec 17;234(5329):410–410. doi: 10.1038/234410a0. [DOI] [PubMed] [Google Scholar]
  11. Goldman R. D., Rebhun L. I. The structure and some properties of the isolated mitotic apparatus. J Cell Sci. 1969 Jan;4(1):179–209. doi: 10.1242/jcs.4.1.179. [DOI] [PubMed] [Google Scholar]
  12. Hinkley R., Telser A. Heavy meromyosin-binding filaments in the mitotic apparatus of mammaliam cells. Exp Cell Res. 1974 May;86(1):161–164. doi: 10.1016/0014-4827(74)90662-4. [DOI] [PubMed] [Google Scholar]
  13. Inoué S., Sato H. Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement. J Gen Physiol. 1967 Jul;50(6 Suppl):259–292. [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kikuchi M., Noda H., Maruyama K. Interaction of actin with H-meromyosin at low ionic strength. J Biochem. 1969 Jun;65(6):945–952. doi: 10.1093/oxfordjournals.jbchem.a129099. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. McIntosh J. R., Cande W. Z., Snyder J. A. Structure and physiology of the mammalian mitotic spindle. Soc Gen Physiol Ser. 1975;30:31–76. [PubMed] [Google Scholar]
  18. McIntosh J. R., Landis S. C. The distribution of spindle microtubules during mitosis in cultured human cells. J Cell Biol. 1971 May 1;49(2):468–497. doi: 10.1083/jcb.49.2.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nicklas R. B. Mitosis. Adv Cell Biol. 1971;2:225–297. doi: 10.1007/978-1-4615-9588-5_5. [DOI] [PubMed] [Google Scholar]
  20. Owaribe K., Hatano S. Inducation of antibody against actin from myxomycete plasmodium and its properties. Biochemistry. 1975 Jul;14(13):3024–3029. doi: 10.1021/bi00684a035. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. RIS H. The anaphase movement of chromosomes in the spermatocytes of the grasshopper. Biol Bull. 1949 Feb;96(1):90–106. [PubMed] [Google Scholar]
  23. Rickards G. K. Prophase chromosome movements in living house cricket spermatocytes and their relationship to prometaphase, anaphase and granule movements. Chromosoma. 1975;49(4):407–455. doi: 10.1007/BF00285133. [DOI] [PubMed] [Google Scholar]
  24. Salmon E. D. Spindle microtubules: thermodynamics of in vivo assembly and role in chromosome movement. Ann N Y Acad Sci. 1975 Jun 30;253:383–406. doi: 10.1111/j.1749-6632.1975.tb19216.x. [DOI] [PubMed] [Google Scholar]
  25. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tawada K., Oosawa F. Effect of the H-meromyosin plus ATP system on F-actin. Biochim Biophys Acta. 1969 May;180(1):199–201. doi: 10.1016/0005-2728(69)90209-6. [DOI] [PubMed] [Google Scholar]
  27. Tilney L. G. Actin filaments in the acrosomal reaction of Limulus sperm. Motion generated by alterations in the packing of the filaments. J Cell Biol. 1975 Feb;64(2):289–310. doi: 10.1083/jcb.64.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tilney L. G., Hatano S., Ishikawa H., Mooseker M. S. The polymerization of actin: its role in the generation of the acrosomal process of certain echinoderm sperm. J Cell Biol. 1973 Oct;59(1):109–126. doi: 10.1083/jcb.59.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]

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