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. 1971 Jun 1;49(3):785–802. doi: 10.1083/jcb.49.3.785

NUCLEOLI OF DIPLOID CELL STRAINS

Their Normal Ultrastructure and the Effects of Toyocamycin and Actinomycin D

Stephanie G Phillips 1, David M Phillips 1
PMCID: PMC2108481  PMID: 5104724

Abstract

Nucleoli of cultured Chinese hamster or mouse cells in early passages had a loosely reticular substructure. Within the reticulum small, irregularly shaped, light fibrillar zones occurred which were contiguous with denser fibrillar zones. These denser zones appeared to be connected in some places to the particulate material which composed the mass of the nucleolus. Generally, electron-transparent spaces separated the particulate zones from the fibrillar areas. Treatment with toyocamycin, an agent which is reported to cause a blockage in the processing of ribosomal RNA, greatly inhibited the accumulation of newly synthesized RNA in the cytoplasm, as monitored by radioautography. Toyocamycin treatment caused the gradual disappearance of the granules from the particulate region of the nucleoli, and resulted ultimately in the nucleoli appearing homogeneously fibrillar. Actinomycin D treatment, which inhibited virtually all RNA synthesis, caused a segregation, and finally a disaggregation, of nucleolar components.

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Selected References

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

  1. BROWN D. D., GURDON J. B. ABSENCE OF RIBOSOMAL RNA SYNTHESIS IN THE ANUCLEOLATE MUTANT OF XENOPUS LAEVIS. Proc Natl Acad Sci U S A. 1964 Jan;51:139–146. doi: 10.1073/pnas.51.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barr H. J., Plaut W. Comparative morphology of nucleolar DNA in Drosophila. J Cell Biol. 1966 Dec;31(3):C17–C22. doi: 10.1083/jcb.31.3.c17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernhard W. Ultrastructural aspects of the normal and pathological nucleolus in mammalian cells. Natl Cancer Inst Monogr. 1966 Dec;23:13–38. [PubMed] [Google Scholar]
  4. Ebstein B. S. The distribution of DNA within the nucleoli of the amphibian oocyte as demonstrated by tritiated actinomycin D radioautography. J Cell Sci. 1969 Jul;5(1):27–44. doi: 10.1242/jcs.5.1.27. [DOI] [PubMed] [Google Scholar]
  5. Geuskens M., Bernhard W. Cytochimie ultrastructurale du nucléole. 3. Action de l'actinomycine D sur le métabolisme du RNA nucléolaire. Exp Cell Res. 1966 Nov-Dec;44(2):579–598. doi: 10.1016/0014-4827(66)90462-9. [DOI] [PubMed] [Google Scholar]
  6. Ghosh S., Lettré R., Ghosh I. On the composition of the nucleolus with special reference to its filamentous structure. Z Zellforsch Mikrosk Anat. 1969;101(2):254–265. doi: 10.1007/BF00335732. [DOI] [PubMed] [Google Scholar]
  7. Hay E. D., Gurdon J. B. Fine structure of the nucleolus in normal and mutant Xenopus embryos. J Cell Sci. 1967 Jun;2(2):151–162. doi: 10.1242/jcs.2.2.151. [DOI] [PubMed] [Google Scholar]
  8. Healy G. M., Parker R. C. Selective nucleolar uptake of alpha-1-acid glycoprotein by mammalian cells in tissue culture. Biochim Biophys Acta. 1967 Nov 28;148(2):556–558. doi: 10.1016/0304-4165(67)90155-9. [DOI] [PubMed] [Google Scholar]
  9. Heine U. Electron microscopic studies on HeLa cells exposed to the antibiotic toyocamycin. Cancer Res. 1969 Oct;29(10):1875–1880. [PubMed] [Google Scholar]
  10. Hillman N., Tasca R. J. Ultrastructural and autoradiographic studies of mouse cleavage stages. Am J Anat. 1969 Oct;126(2):151–173. doi: 10.1002/aja.1001260203. [DOI] [PubMed] [Google Scholar]
  11. Hyde B. B. Changes in nucleolar ultrastructure associated with differentiation in the root apex. Natl Cancer Inst Monogr. 1966 Dec;23:39–52. [PubMed] [Google Scholar]
  12. Karasaki S. The ultrastructure and RNA metabolism of nucleoli in early sea urchin embryos. Exp Cell Res. 1968 Sep;52(1):13–26. doi: 10.1016/0014-4827(68)90543-0. [DOI] [PubMed] [Google Scholar]
  13. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. La Cour L. F., Wells B. The loops and ultrastructure of the nucleolus of Ipheion uniflorum. Z Zellforsch Mikrosk Anat. 1967;82(1):25–45. doi: 10.1007/BF00326099. [DOI] [PubMed] [Google Scholar]
  15. Lane N. J. Spheroidal and ring nucleoli in amphibian oocytes. Patterns of uridine incorporation and fine structural features. J Cell Biol. 1967 Nov;35(2):421–434. doi: 10.1083/jcb.35.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Macgregor H. C. Pattern of incorporation of (3H)uridine into RNA of amphibian oocyte nucleoli. J Cell Sci. 1967 Jun;2(2):145–150. doi: 10.1242/jcs.2.2.145. [DOI] [PubMed] [Google Scholar]
  17. Miller O. L., Jr Structure and composition of peripheral nucleoli of salamander oocytes. Natl Cancer Inst Monogr. 1966 Dec;23:53–66. [PubMed] [Google Scholar]
  18. Mundell R. D. The occurrence of ribosomal proteins in nucleoli of starfish oocytes. Biochem Biophys Res Commun. 1967 Jul 10;28(1):117–122. doi: 10.1016/0006-291x(67)90415-9. [DOI] [PubMed] [Google Scholar]
  19. Muramatsu M., Hodnett J. L., Steele W. J., Busch H. Synthesis of 28-S RNA in the nucleolus. Biochim Biophys Acta. 1966 Jul 20;123(1):116–125. doi: 10.1016/0005-2787(66)90164-x. [DOI] [PubMed] [Google Scholar]
  20. PELLING C. RIBONUKLEINSAEURE-SYNTHESE DER RIESENCHROMOSOMEN. AUTORADIOGRAPHISCHE UNTERSUCHUNGEN AN CHIRONOMUS TENTANS. Chromosoma. 1964 Apr 1;15:71–122. doi: 10.1007/BF00326915. [DOI] [PubMed] [Google Scholar]
  21. Perry R. P., Cheng T. Y., Freed J. J., Greenberg J. R., Kelley D. E., Tartof K. D. Evolution of the transcription unit of ribosomal RNA. Proc Natl Acad Sci U S A. 1970 Mar;65(3):609–616. doi: 10.1073/pnas.65.3.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Phillips D. M., Phillips S. G. Distinctive characteristics of nucleoli of two established cell lines. J Cell Biol. 1971 Jun;49(3):803–815. doi: 10.1083/jcb.49.3.803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Phillips S. G., Phillips D. M. Sites of nucleolus production in cultured Chinese hamster cells. J Cell Biol. 1969 Jan;40(1):248–268. doi: 10.1083/jcb.40.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. RITOSSA F. M., SPIEGELMAN S. LOCALIZATION OF DNA COMPLEMENTARY TO RIBOSOMAL RNA IN THE NUCLEOLUS ORGANIZER REGION OF DROSOPHILA MELANOGASTER. Proc Natl Acad Sci U S A. 1965 Apr;53:737–745. doi: 10.1073/pnas.53.4.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. STEVENS B. J. THE EFFECT OF ACTINOMYCIN D ON NUCLEOLAR AND NUCLEAR FINE STRUCTURE IN THE SALIVARY GLAND CELL OF HIRONOMUS THUMMI. J Ultrastruct Res. 1964 Oct;11:329–353. doi: 10.1016/s0022-5320(64)90037-1. [DOI] [PubMed] [Google Scholar]
  26. Svoboda D., Racela A., Higginson J. Variations in ultrastructural nuclear changes in hepatocarcinogenesis. Biochem Pharmacol. 1967 Apr;16(4):651–657. doi: 10.1016/0006-2952(67)90077-9. [DOI] [PubMed] [Google Scholar]
  27. Swift H., Stevens B. J. Nucleolar-chromosomal interaction in microspores of maize. Natl Cancer Inst Monogr. 1966 Dec;23:145–166. [PubMed] [Google Scholar]
  28. Tavitian A., Uretsky S. C., Acs G. Selective inhibition of ribosomal RNA synthesis in mammalian cells. Biochim Biophys Acta. 1968 Mar 18;157(1):33–43. doi: 10.1016/0005-2787(68)90261-x. [DOI] [PubMed] [Google Scholar]
  29. Tavitian A., Uretsky S. C., Acs G. The effect of toyocamycin on cellular RNA synthesis. Biochim Biophys Acta. 1969 Mar 18;179(1):50–57. doi: 10.1016/0005-2787(69)90121-x. [DOI] [PubMed] [Google Scholar]
  30. VENABLE J. H., COGGESHALL R. A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY. J Cell Biol. 1965 May;25:407–408. doi: 10.1083/jcb.25.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vincent W. S., Baltus E., Lovlie A., Mundell R. E. Proteins and nucleic acids of starfish oocyte nucleoli and ribosomes. Natl Cancer Inst Monogr. 1966 Dec;23:235–253. [PubMed] [Google Scholar]
  32. Weinberg R. A., Loening U., Willems M., Penman S. Acrylamide gel electrophoresis of HeLa cell nucleolar RNA. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1088–1095. doi: 10.1073/pnas.58.3.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weinberg R. A., Penman S. Small molecular weight monodisperse nuclear RNA. J Mol Biol. 1968 Dec;38(3):289–304. doi: 10.1016/0022-2836(68)90387-2. [DOI] [PubMed] [Google Scholar]
  34. von Gaudecker B. RNA synthesis in the nucleolus of Chironomus Thummi, as studied by high resolution autoradiography. Z Zellforsch Mikrosk Anat. 1967;82(4):536–557. doi: 10.1007/BF00337121. [DOI] [PubMed] [Google Scholar]

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