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Isolation and Characterization of Mitoplasts

  • Potu N. Rao
  • Prasad S. Sunkara
  • Abdullatif A. Al-Bader

Abstract

The discovery by Carter (1967) that cytochalasin B (CB), a metabolite produced by the fungus Helminthosporium dematiodeum, causes nuclear extrusion and spontaneous enucleation in a small proportion of treated cells has led to the development of a number of techniques to study nucleocyto-plasmic interactions, cytoplasmic inheritance, and the creation of reconstituted cells. CB has a profound effect on the morphology of mammalian cells in culture and is best known for blocking cytokinesis without affecting the karyokinesis or the nuclear division that usually leads to the formation of binucleate cells (Carter, 1967; Ladda and Estensen, 1970). However, the unique feature of CB, its induction of spontaneous nuclear extrusion, has been exploited by the cell biologist to the maximum extent.

Keywords

Mitotic Cell Interphase Nucleus Mitotic Chromosome Interphase Cell Ficoll Gradient 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Brinkley, B. R., and Rao, P. N., 1973, Nitrous oxide: Effects on the mitotic apparatus and chromosome movement in HeLa cells, J. Cell Biol. 58:96–106.PubMedCrossRefGoogle Scholar
  2. Carter, S. B., 1967, Effect of cytochalasins on mammalian cells, Nature 213:261–264.PubMedCrossRefGoogle Scholar
  3. Eigsti, O. J., and Dustin, P., 1955, Colchicine in Agriculture, Medicine, Biology and Chemistry, Iowa State University Press, Ames, Iowa.Google Scholar
  4. Gopalakrishnan, T. V., and Thompson, E. B., 1975, A method for enucleating cultured mammalian cells, Exp. Cell Res. 96:435–439.PubMedCrossRefGoogle Scholar
  5. Johnson, R. T., and Rao, P. N., 1970, Mammalian cell fusion. II. Induction of premature chromosome condensation in interphase nuclei, Nature 226:717–722.PubMedCrossRefGoogle Scholar
  6. Ladda, R. L., and Estensen, R. D., 1970, Introduction of a heterologous nucleus into enucleated cytoplasms of cultured mouse L-cells, Proc. Natl. Acad. Sci. USA 67:1528–1533.PubMedCrossRefGoogle Scholar
  7. Mans, R. J., and Novelli, G. D., 1961, Measurement of the incorporation of radio-active amino acids into protein by a filter-paper disk method, Arch. Biochem. Biophys. 94:48–53.CrossRefGoogle Scholar
  8. Prescott, D. M., 1976, The cell cycle and the control of cellular reproduction, Adv. Genet. 18:99–177.PubMedCrossRefGoogle Scholar
  9. Prescott, D. M., Myerson, D., and Wallace, J., 1971, Enucleation of mammalian cells with cytochalasin B, Exp. Cell Res. 71:480–485.CrossRefGoogle Scholar
  10. Rao, P. N., 1968, Mitotic synchrony in mammalian cells treated with nitrous oxide at high pressure, Science 160:774–776.PubMedCrossRefGoogle Scholar
  11. Rao, P. N., and Engelberg, J., 1966, Effects of temperature on the mitotic cycle of normal and synchronized mammalian cells, in: Cell Synchrony — Biosynthetic Regulation, (I. L. Cameron and G. M. Padilla, eds.), Academic Press, New York, pp. 332–352.Google Scholar
  12. Rao, P. N., and Johnson, R. T., 1972, Cell fusion and its application to studies on the regulation of cell cycle, in: Methods in Cell Physiology, Volume V (D. M. Prescott, ed.), Academic Press, New York, pp. 75–126.Google Scholar
  13. Rao, P. N., and Johnson, R. T., 1974, Regulation of cell cycle in hybrid cells, in: Control of Proliferation in Animal Cells, Cold Spring Harbor Conferences on Cell Proliferation, Volume 1, Cold Spring Harbor Laboratories, Cold Spring Harbor, pp. 785–800.Google Scholar
  14. Sunkara, P. S., Al-Bader, A. A., and Rao, P. N., 1977, Mitoplasts: Mitotic cells minus the chromosomes, Exp. Cell Res. 107:444–448.PubMedCrossRefGoogle Scholar
  15. Sunkara, P. S., Al-Bader, A. A., Riker, M. A., and Rao, P. N., 1980, Induction of prematurely condensed chromosomes by mitoplasts, Cell Biol. Internatl. Rep. 4:1025–1029.CrossRefGoogle Scholar
  16. Tabor, C. W., 1962, Stabilization of protoplasts and spheroplasts by spermine and other polyamines, J. Bacteriol. 83:1101–1111.PubMedGoogle Scholar
  17. Veomett, G., Shay, J., Hough, P. V. C., and Prescott, D. M., 1976, Large-scale enucleation of mammalian cells, in: Methods in Cell Biology, Volume XIII (D. M. Prescott, ed.), Academic Press, New York, pp. 1–14.Google Scholar
  18. Wigler, M. H., and Weinstein, I. B., 1975, A preparative method for obtaining enucleated mammalian cells, Biochem. Biophys. Res. Commun. 63:669–674.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Potu N. Rao
    • 1
  • Prasad S. Sunkara
    • 2
  • Abdullatif A. Al-Bader
    • 3
  1. 1.Department of Developmental Therapeutics, M. D. Anderson Hospital and Tumor InstituteUniversity of Texas System Cancer CenterHoustonUSA
  2. 2.Merrell Research CenterCincinnatiUSA
  3. 3.Department of PathologyKuwait UniversityKuwait

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