Advertisement

Chromosoma

, Volume 53, Issue 1, pp 37–50 | Cite as

Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining

  • Carll Goodpasture
  • Stephen E. Bloom
Article

Abstract

A simple ammoniacal silver staining procedure, designated Ag-AS, differentially stains the chromosomal locations of ribosomal DNA in certain mammalian species. This was critically demonstrated by Ag-AS staining of the nucleolus organizer regions in karyotypes of the same species and cell lines used for locating the ribosomal cistrons by DNA/RNA in situ hybridization. With Ag-AS, silver stained NORs (Ag-NORs) are visualized as black spherical bodies on yellow-brown chromosome arms. Ag-NORs were visualized throughout mitosis at the secondary constrictions in the rat kangaroo, Seba's fruit bat, Indian muntjac, and Rhesus monkey. The Chinese hamster and cattle have telomeric Ag-NORs, the mouse subcentromeric Ag-NORs, and the field vole Ag-NORs as minute short arms or chromosomal satellites. Ag-NORs occur at both secondary constrictions and at telomeres in the cotton rat. Variability in Ag-NOR pattern included differences in the number of Ag-NORs per cell within a cell population, size of Ag-NORs among chromosomes of a complement, and presence of Ag-NOR on particular chromosomes in two cell lines of the Chinese hamster. The available cytochemical data suggest that the Ag-AS reaction stains chromosomal proteins at the NOR rather than the rDNA itself.

Keywords

Silver Staining Organizer Region Nucleolar Organizer Region Nucleolar Organizer Secondary Constriction 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bartalos, M., Rainer, J. O.: Human chromosome mapping with an ammoniacal silver staining procedure. Acta Genet. Med. Gemellol. 21, 139–142 (1972)Google Scholar
  2. Black, M. M., Ansley, H. R.: Histone staining with ammoniacal silver. Science 143, 693–695 (1964)Google Scholar
  3. Black, M. M., Ansley, H. R.: Histone specificity revealed by ammoniacal silver staining. J. Histochem. Cytochem. 14, 177–181 (1966)Google Scholar
  4. Black, M. M., Speer, F. D., Lillick, L. C.: Acid-extractable nuclear proteins of cancer cells. I. Staining with ammoniacal silver. J. nat. Cancer Inst. 25, 967–977 (1960)Google Scholar
  5. Bloom, S. E., Buss, E. G.: Ammoniacal silver staining of embryonic chicken cells and chromosomes. Poultry Sci. 48, 1114–1116 (1969)Google Scholar
  6. Das, N. K.: Demonstration of a non-RNA nucleolar fraction by silver staining. Exp. Cell Res. 26, 428–431 (1961)Google Scholar
  7. Denton, T. E., Howell, W. M., Barrett, J. V.: Human nucleolar organizer chromosomes: satellite associations (in press, 1975)Google Scholar
  8. Evans, H. J., Buckland, R. A., Pardue, M. L.: Location of the genes coding for 18s and 28s ribosomal RNA in the human genome. Chromosoma (Berl.) 48, 405–426 (1974)Google Scholar
  9. Funaki, K., Matsui, S., Sasaki, M.: Location of nucleolar organizers in animal and plant chromosomes by means of an improved N-banding technique. Chromosoma (Berl.) 49, 357–370 (1975)Google Scholar
  10. Henderson, A. S., Eicher, E. M., Yu, M. T., Atwood, K. C.: The chromosomal location of ribosomal DNA in the mouse. Chromosoma (Berl.) 49, 155–160 (1974a)Google Scholar
  11. Henderson, A. S., Warburton, D., Atwood, K. C.: Location of rDNA in the human chromosome complement. Proc. nat. Acad. Sci. (Wash.) 69, 3394–3398 (1972)Google Scholar
  12. Henderson, A. S., Warburton, D., Atwood, K. C.: Localization of rDNA in the chromosome complement of the Rhesus (Macaca mulatta). Chromosoma (Berl.) 44, 367–370 (1974b)Google Scholar
  13. Howell, W. M., Denton, T. E.: An ammoniacal-silver stain technique specific for satellite III DNA regions on human chromosomes. Experientia (Basel) 30, 1364–1366 (1974)Google Scholar
  14. Howell, W. M., Denton, T. E., Diamond, J. R.: Differential staining of the satellite regions of human acrocentric chromosomes. Experientia (Basel) 31, 260–262 (1975)Google Scholar
  15. Hsu, T. C.: Longitudinal differentiation of chromosomes. Ann. Rev. Genet. 7, 153–175 (1973)Google Scholar
  16. Hsu, T. C., Spirito, S. E., Pardue, M. L.: Distribution of 18+28S ribosomal genes in mammalian genomes. Chromosoma (Berl.) 53, 25–36 (1975)Google Scholar
  17. Hsu, T. C., Zenzes, M. T.: Mammalian chromosomes in vitro. XVII. Idiogram of the Chinese hamster. J. nat. Cancer Inst. 32, 857–869 (1964)Google Scholar
  18. Kato, H., Tsuchiya, K., Yosida, T. H.: Constitutive heterochromatin of Indian muntjac chromosomes revealed by DNase treatment and a C-banding technique. Canad. J. Genet. Cytol. 16, 273–280 (1974)Google Scholar
  19. Krunit, D. M., Shafit, B. R., Maio, J. J.: Multiple satellite deoxyribonucleic acids in the calf and their relation to the sex chromosomes. J. molec. Biol. 81, 273–284 (1973)Google Scholar
  20. Matsui, S.: Structural proteins associated with ribosomal cistrons in Xenopus laevis chromosomes. Exp. Cell Res. 88, 88–94 (1974)Google Scholar
  21. Matsui, S., Sasaki, M.: Differential staining of nucleolus organizers in mammalian chromosomes. Nature (Lond.) 246, 148–150 (1973)Google Scholar
  22. Pardue, M. L., Hsu, T. C.: Locations of 18s and 28s ribosomal genes on the chromosomes of the Indian muntjac. J. Cell Biol. 64, 251–254 (1975)Google Scholar
  23. Rothfels, K. H., Siminovitch, L.: The chromosome complement of the Rhesus monkey (Macaca mulatta) determined in kidney cells cultivated in vitro. Chromosoma (Berl.) 9, 163–175 (1958)Google Scholar
  24. Sinclair, J. H., Brown, D. D.: Retention of common nucleotide sequences in the ribosomal deoxyribonucleic acid of eukaryotes and some of their physical characteristics. Biochemistry 10, 2761–2769 (1971)Google Scholar
  25. Utsumi, S., Takehisa, S.: Heterochromatin differentiation in Trillium kamtschaticum by ammoniacal silver reaction. Exp. Cell Res. 86, 398–401 (1974)Google Scholar
  26. Wimber, D. E., Steffensen, D. M.: Localization of gene function. Ann. Rev. Genet. 7, 205–223 (1973)Google Scholar
  27. Zimmerman, E. G., Lee, M. R.: Variation in chromosomes of the cotton rat, Sigmodon hispidus. Chromosoma (Berl.) 24, 243–250 (1968)Google Scholar
  28. Zimmerman, E. G., Sihronen, D. A.: Chromosomal banding pattern and idiogram of the cotton rat, Sigmodon arizonae (Rodentia, Muridae). Chromosoma (Berl.) 41, 85–91 (1973)Google Scholar

Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • Carll Goodpasture
    • 1
  • Stephen E. Bloom
    • 1
  1. 1.Department of BiologyThe University of Texas System Cancer Center M. D. Anderson Hospital and Tumor InstituteHoustonUSA

Personalised recommendations