Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1971 Jun 1;49(3):830–847. doi: 10.1083/jcb.49.3.830

ULTRASTRUCTURE OF THE NUCLEOLUS DURING THE CHINESE HAMSTER CELL CYCLE

J S Noel 1, W C Dewey 1, J H Abel Jr 1, R P Thompson 1
PMCID: PMC2108503  PMID: 4933472

Abstract

Changes in the structure of the nucleolus during the cell cycle of the Chinese hamster cell in vitro were studied. Quantitative electron microscopic techniques were used to establish the size and volume changes in nucleolar structures. In mitosis, nucleolar remnants, "persistent nucleoli," consisting predominantly of ribosome-like granular material, and a granular coating on the chromosomes were observed. Persistent nucleoli were also observed in some daughter nuclei as they were leaving telophase and entering G1. During very early G1, a dense, fibrous material characteristic of interphase nucleoli was noted in the nucleoplasm of the cells. As the cells progressed through G1, a granular component appeared which was intimately associated with the fibrous material. By the middle of G1, complete, mature nucleoli were present. The nucleolar volume enlarged by a factor of two from the beginning of G1 to the middle of S primarily due to the accumulation of the granular component. During the G2 period, there was a dissolution or breakdown of the nucleolus prior to the entry of the cells into mitosis. Correlations between the quantitative aspects of this study and biochemical and cytochemical data available in the literature suggest the following: nucleolar reformation following division results from the activation of the nucleolar organizer regions which transcribe for RNA first appearing in association with protein as a fibrous component (45S RNA) and then later as a granular component (28S and 32S RNA).

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. 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]
  2. Brinkley B. R. The fine structure of the nucleolus in mitotic divisions of Chinese hamster cells in vitro. J Cell Biol. 1965 Nov;27(2):411–422. doi: 10.1083/jcb.27.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown D. D., Gurdon J. B. Size distribution and stability of DNA-like RNA synthesized during development of anucleolate embryos of Xenopus laevis. J Mol Biol. 1966 Aug;19(2):399–422. doi: 10.1016/s0022-2836(66)80013-x. [DOI] [PubMed] [Google Scholar]
  4. Dewey W. C., Humphrey R. M., Sedita B. A. Cell cycle kinetics and radiation-induced chromosomal aberrations studied with C14 and H3 labels. Biophys J. 1966 May;6(3):247–260. doi: 10.1016/S0006-3495(66)86654-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dewey W. C., Miller H. H. X-ray induction of chromatid exchanges in mitotic and G1 Chinese hamster cells pretreated with Colcemid. Exp Cell Res. 1969 Sep;57(1):63–70. doi: 10.1016/0014-4827(69)90366-8. [DOI] [PubMed] [Google Scholar]
  6. GIRARD M., LATHAM H., PENMAN S., DARNELL J. E. ENTRANCE OF NEWLY FORMED MESSENGER RNA AND RIBOSOMES INTO HELA CELL CYTOPLASM. J Mol Biol. 1965 Feb;11:187–201. doi: 10.1016/s0022-2836(65)80050-x. [DOI] [PubMed] [Google Scholar]
  7. GIRARD M., PENMAN S., DARNELL J. E. THE EFFECT OF ACTINOMYCIN ON RIBOSOME FORMATION IN HELA CELLS. Proc Natl Acad Sci U S A. 1964 Feb;51:205–211. doi: 10.1073/pnas.51.2.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GRANBOULAN N., GRANBOULAN P. CYTOCHIMIE ULTRASTRUCTURALE DU NUCL'EOLE. I. MISE EN 'EVIDENCE DE CHROMATINE 'A L'INT'ERIEUR DU NUCL'EOLE. Exp Cell Res. 1964 Mar;34:71–87. doi: 10.1016/0014-4827(64)90184-3. [DOI] [PubMed] [Google Scholar]
  9. GRANBOULAN N., GRANBOULAN P. CYTOCHIMIE ULTRASTRUCTURALE DU NUCL'EOLE. II. ETUDE DES SITES DE SYNTH'ESE DU RNA DANS LE NUCL'EOLE ET LE NOYAU. Exp Cell Res. 1965 Jun;38:604–619. doi: 10.1016/0014-4827(65)90384-8. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. HARRIS H. The initiation of deoxyribonucleic acid synthesis in the connective-tissue cell, with some observations on the function of the nucleolus. Biochem J. 1959 May;72(1):54–60. doi: 10.1042/bj0720054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Heneen W. K., Nichols W. W. Persistence of nucleoli in short term and long term cell cultures and in direct bone marrow preparations in mammalian materials. J Cell Biol. 1966 Dec;31(3):543–561. doi: 10.1083/jcb.31.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hsu T. C., Arrighi F. E., Klevecz R. R., Brinkley B. R. The nucleoli in mitotic divisions of mammalian cells in vitro. J Cell Biol. 1965 Aug;26(2):539–553. doi: 10.1083/jcb.26.2.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hsu T. C., Brinkley B. R., Arrighi F. E. The structure and behavior of the nucleolus organizer in mammalian cells. Chromosoma. 1967;23(2):137–153. doi: 10.1007/BF00331109. [DOI] [PubMed] [Google Scholar]
  16. Hyde B. B., Sankaranarayanan K., Birnstiel M. L. Observations on fine structure in pea nucleoli in situ and isolated. J Ultrastruct Res. 1965 Jun;12(5):652–667. doi: 10.1016/s0022-5320(65)80054-5. [DOI] [PubMed] [Google Scholar]
  17. JONES O. P. Paramitotic granulation and ribosome bodies in erythroblasts. J Ultrastruct Res. 1962 Oct;7:308–315. doi: 10.1016/s0022-5320(62)90026-6. [DOI] [PubMed] [Google Scholar]
  18. Jacob J. An electron microscope autoradiographic study of the site of initial synthesis of RNA in the nucleolus of Smittia. Exp Cell Res. 1967 Nov;48(2):276–282. doi: 10.1016/0014-4827(67)90353-9. [DOI] [PubMed] [Google Scholar]
  19. LAFONTAINE J. G., CHOUINARD L. A. A correlated light and electron microscope study of the nucleolar material during mitosis in Vicia faba. J Cell Biol. 1963 Apr;17:167–201. doi: 10.1083/jcb.17.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. LOUD A. V., BARANY W. C., PACK B. A. QUANTITATIVE EVALUATION OF CYTOPLASMIC STRUCTURES IN ELECTRON MICROGRAPHS. Lab Invest. 1965 Jun;14:996–1008. [PubMed] [Google Scholar]
  21. 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]
  22. Liau M. C., Perry R. P. Ribosome precursor particles in nucleoli. J Cell Biol. 1969 Jul;42(1):272–283. doi: 10.1083/jcb.42.1.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lord A., Lafontaine J. G. The organization of the nucleolus in meristematic plant cells. A cytochemical study. J Cell Biol. 1969 Mar;40(3):633–647. doi: 10.1083/jcb.40.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MARINOZZI V. CYTOCHIMIE ULTRASTRUCTURALE DU NUCL'EOLE--RNA ET PROT'EINES INTRANUCL'EOLAIRES. J Ultrastruct Res. 1964 Jun;10:433–456. doi: 10.1016/s0022-5320(64)80021-6. [DOI] [PubMed] [Google Scholar]
  25. Miller O. L., Jr, Beatty B. R. Visualization of nucleolar genes. Science. 1969 May 23;164(3882):955–957. doi: 10.1126/science.164.3882.955. [DOI] [PubMed] [Google Scholar]
  26. Murray R. G., Murray A. S., Pizzo A. The fine structure of mitosis in rat thymic lymphocytes. J Cell Biol. 1965 Aug;26(2):601–619. doi: 10.1083/jcb.26.2.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. PALADE G. E. A study of fixation for electron microscopy. J Exp Med. 1952 Mar;95(3):285–298. doi: 10.1084/jem.95.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. PERRY R. P. ROLE OF THE NUCLEOLUS IN RIBONUCLEIC ACID METABOLISM AND OTHER CELLULAR PROCESSES. Natl Cancer Inst Monogr. 1964 May;14:73–89. [PubMed] [Google Scholar]
  29. PRESCOTT D. M., BENDER M. A. Synthesis of RNA and protein during mitosis in mammalian tissue culture cells. Exp Cell Res. 1962 Mar;26:260–268. doi: 10.1016/0014-4827(62)90176-3. [DOI] [PubMed] [Google Scholar]
  30. Penman S., Smith I., Holtzman E., Greenberg H. RNA metabolism in the HeLa cell nucleus and nucleolus. Natl Cancer Inst Monogr. 1966 Dec;23:489–512. [PubMed] [Google Scholar]
  31. Perry R. P., Kelley D. E. Buoyant densities of cytoplasmic ribonucleoprotein particles of mammalian cells: distinctive character of ribosome subunits and the rapidly labeled components. J Mol Biol. 1966 Apr;16(2):255–268. doi: 10.1016/s0022-2836(66)80171-7. [DOI] [PubMed] [Google Scholar]
  32. Perry R. P. On ribosome biogenesis. Natl Cancer Inst Monogr. 1966 Dec;23:527–545. [PubMed] [Google Scholar]
  33. Perry R. P. The nucleolus and the synthesis of ribosomes. Natl Cancer Inst Monogr. 1965 Dec;18:325–340. [PubMed] [Google Scholar]
  34. 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]
  35. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. ROBBINS E., GONATAS N. K. THE ULTRASTRUCTURE OF A MAMMALIAN CELL DURING THE MITOTIC CYCLE. J Cell Biol. 1964 Jun;21:429–463. doi: 10.1083/jcb.21.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. SCHOEFL G. I. THE EFFECT OF ACTINOMYCIN D ON THE FINE STRUCTURE OF THE NUCLEOLUS. J Ultrastruct Res. 1964 Apr;10:224–243. doi: 10.1016/s0022-5320(64)80007-1. [DOI] [PubMed] [Google Scholar]
  38. SMETANA K., BUSCH H. STUDIES ON THE ULTRASTRUCTURE OF THE NUCLEOLI OF THE WALKER TUMOR AND RAT LIVER. Cancer Res. 1964 May;24:537–557. [PubMed] [Google Scholar]
  39. STEVENS B. J. THE FINE STRUCTURE OF THE NUCLEOLUS DURING MITOSIS IN THE GRASSHOPPER NEUROBLAST CELL. J Cell Biol. 1965 Mar;24:349–368. doi: 10.1083/jcb.24.3.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shinozuka H., Reid I. M., Shull K. H., Liang H., Farber E. Dynamics of liver cell injury and repair. I. Spontaneous reformation of the nucleolus and polyribosomes in the presence of extensive cytoplasmic damage induced by ethionine. Lab Invest. 1970 Sep;23(3):253–267. [PubMed] [Google Scholar]
  41. TAYLOR J. H. Nucleic acid synthesis in relation to the cell division cycle. Ann N Y Acad Sci. 1960 Oct 7;90:409–421. doi: 10.1111/j.1749-6632.1960.tb23259.x. [DOI] [PubMed] [Google Scholar]
  42. Terzakis J. A. The nucleolar channel system of human endometrium. J Cell Biol. 1965 Nov;27(2):293–304. doi: 10.1083/jcb.27.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tobey R. A., Anderson E. C., Petersen D. F. Properties of mitotic cells prepared by mechanically shaking monolayer cultures of Chinese hamster cells. J Cell Physiol. 1967 Aug;70(1):63–68. doi: 10.1002/jcp.1040700109. [DOI] [PubMed] [Google Scholar]
  44. Unuma T., Arendell J. P., Busch H. High resolution autoradiographic studies of the uptake of 3H-5-uridine into condensed and dispersed chromatin of nuclei and granular and fibrillar components of nucleoli of Novikoff hepatoma ascites cells. Exp Cell Res. 1968 Oct;52(2):429–438. doi: 10.1016/0014-4827(68)90484-9. [DOI] [PubMed] [Google Scholar]
  45. Unuma T., Floyd L. R., Busch H. Selective removal of the perinucleolar nucleolus-associated chromatin from the isolated nucleoli of livers of thioacetamide-treated rats. Exp Cell Res. 1968 Sep;52(1):101–111. doi: 10.1016/0014-4827(68)90550-8. [DOI] [PubMed] [Google Scholar]
  46. Willems M., Penman M., Penman S. The regulation of RNA synthesis and processing in the nucleolus during inhibition of protein synthesis. J Cell Biol. 1969 Apr;41(1):177–187. doi: 10.1083/jcb.41.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yasuzumi G., Sugihara R. The fine structure of nuclei as revealed by electron microscopy. II. The progress of nucleolus reconstitution in Ehrlich ascites tumor cell nuclei. Exp Cell Res. 1965 Oct;40(1):45–55. doi: 10.1016/0014-4827(65)90288-0. [DOI] [PubMed] [Google Scholar]
  48. Zobel C. R., Beer M. The use of heavy metal salts as electron stains. Int Rev Cytol. 1965;18:363–400. doi: 10.1016/s0074-7696(08)60558-6. [DOI] [PubMed] [Google Scholar]
  49. 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]

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

RESOURCES