Identification of Nucleolus Organizer Regions of Korean Cattle Chromosomes by AgNOR Staining

AgNOR 염색법에 의한 한우 염색체의 Nucleolus Organizer Regions 양상 분석

  • Jung , W. (Department of Animal Science and Biotechnology & RAIRC, Jinju National University) ;
  • Sohn, S.H. (Department of Animal Science and Biotechnology & RAIRC, Jinju National University)
  • 정원 (진주산업대학교 동물생명과학과 . 동물생명산업지역협력연구센터) ;
  • 손시환 (진주산업대학교 동물생명과학과 . 동물생명산업지역협력연구센터)
  • Published : 2003.10.31


Nucleolus Organizer Regions (NORs) are the specific chromosome sites where ribosomal genes are located and highly expressed. We have applied the AgNOR staining to identify the distribution of NORs in the chromosomes of Korean Cattle. We have also studied the NORs pattern on the cells originated from different breeds, tissues and sex. Peripheral blood from forty-four Korean Cattle and Holstein was cultured for chromosme preparation. The fibroblast culture from biopsied ear skins was also conducted for chromosome analysis. The distribution of NORs was analyzed by sequential Ag staining and G-banding on metaphases of the cells. In Korean Cattle, the NORs are localized on the telomeres of the five chromosome pairs number 2, 3, 4, 11 and 28. The number of NORs per metaphase ranged from 2 to 10 giving a mean value of 5.6. The number of NORs per cell varied among individuals and cells within same individual. The size of NORs also differed in NO-chromosomes. The number of NORs was significantly different between Korean Cattle and Holstein, fibroblasts and lymphocytes, and male and female. However, the distribution and frequency of NORs were similar among the cells regardless of breeds, tissues, and sex.


Korean Cattle;AgNOR;NORs;G-banding;Chromosomes


  1. Mayr, B. and Czaker, R. 1981. Variable position of nucleolus organizer regions in Bovidae. Experientia 37, 564-565.
  2. Bloom, S. E. and Goodpasture, C. 1976. An improved technique for selective silver staining of nucleolar organizer regions in human chromosomes. Humangenetik 34:1999-206.
  3. Bloom, S. E., Briles, W. E., Briles, R. W., Delany, M. E. and Dietert, R. R. 1987. Chromosomal localization of the major histocompatibility (B) complex (MHC) and its expression in chickens aneuploid for the major deoxyribonucleic acid microchromosome. Poultry Sci. 66:782-789.
  4. Derenzini, M. and Ploton, D. 1991. Interphase nucleolar organizer regions in cancer cells. Int. Rev. Exp. Pathol 32:150-192.
  5. Di Berardino, D., Arrighi, F. E. and Kieffer, N. M. 1979. Nucleolus organizer regions in two species of Bovidae. J. Heredity 70:47-50.
  6. Di Berardino, D., Iannuzzi, L., Bettini, T. M. and Matassino, D. 1981. Ag-NORs variation banding homologies in two species of bovidae: Bubalus bubalis and Bos taurus. Can. J. Genet. Cytol. 23:89-99.
  7. Di Meo, G. P., Iannuzzi, L., Ferrara, L. and Rubino, R. 1991. Identification of nucleolus organizer chromosomes in goat. Caryologia 44:309-316.
  8. Di Meo, G. P., Iannuzzi, L., Perucatti, A. and Ferrara, L. 1993. Identification of nucleolus organizer chromosomes in sheep. Cytobios 75:183-190.
  9. Diamond, J. R., Dunn, H. O. and Howell, W. M. 1975. Centromeric and telomeric staining regions in the chromosomes of cattle(Bos taurus). Cytogent. Cell Genet. 15:332-337.
  10. Evans, H. J., Buckland, R. A. and Pardue, M. L. 1974. Location of the genes coding 18S and 28S ribosomal RNA in the human genome. Chromosoma 48:405-426.
  11. Gall, J. G. and Pardue, M. L. 1969. Molecular hybridization of radioactive DNA to the DNA of cytological preparation. Proc. Natl. Acad. Sci. USA 64:600-604.
  12. Gallagher Jr., D. S., Davis, S. K., De Donato, M., Burzlaff, J. D., Womack, J. E., Taylor, J. F. and Kumamoto, A. T. 1999. A molecular cytogenetic analysis of the tribe Bovini with an emphasis on sex chromosome morphology and NOR distribution. Chromosome Res. 7:481-492.
  13. Goodpasture, C. and Bloom, S. E. 1975. Visualisation of nucleolus organizer regions in mammalian chromosomes using silver staining. Chromosoma 55:37-50.
  14. Hartung, M., Mirre, C. and Stahl, A. 1979. Nucleolar organizers in human oocytes at meiotic prophase I, studied by the silver-NOR method and electron microscopy. Hum. Genet. 52:295-308.
  15. Henderson, A. S., Warburton, D. and Atwood, K. C. 1972. Location of ribosomal DNA in the human chromosome complement. PNAS 69:3394-3398.
  16. Hendeson, L. M. and Bruere, A. N. 1979. Conservation of nucleolus organizer regions during evolution in sheep, goat, cattle and aoudad. Can. J. Genet. Cytol. 21:1-8.
  17. Howell, W. M. 1977. Visualisation of ribosomal gene activity: silver stains proteins associated with rRNA transcribed from oocyte chromosomes. Chromosoma 62:361-367.
  18. Howell, W. M. and Black, D. A. 1980. Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a 1 step method. Experiantia 36:1014-1015.
  19. Hubbel, H. R. 1985. Silver staining as an indicator active ribosomal genes. Stain Technol. 60:285-294.
  20. Iannuzzi, L., Di Meo, G. P. and Perucatti, A. 1996. Identification of nucleolus organizer chromosomes and frequency of active NORs in river buffalo(Bubalus bubalis L.). Caryologia 49:27-24.
  21. ISCNDB 2000. 2001. International system for chromosome nomenclature of domestic Bovids. Cytogenet. Cell Genet. 92:283-299.
  22. Mayr, B., Gruber, K., Brem, G. and Mayrhofer, G. 1989. Genetic studies on nucleolus organizer regions (NORs) in cattle. Genet. Res. 53:111-118.
  23. Mayr, B., Schleger, W. and Auer, H. 1987. Frequency of Ag-stained nucleolus organizer regions in the chromosomes of cattle. J. Heredity 78:206-207.
  24. Mellink, C. H. M., Bosma, A. A. and De Haan, N. A. 1994. Variation in size of Ag-NORs and fluorescent rDNA in situ hybridization signals in six breeds of domesitic pig. Hereditas 120:141-149.
  25. Mellink, C. H. M., Bosma, A. A., De Haan, N. A. and Macdonald, A. A. 1992. Numerical variation of nucleolar organizer regions after silver staining in domestic and wild Suidae (Mammalia). Animal Genetics 23:231-239.
  26. Miller, D. A., Tantravahi, R., Dev, V. G. and Miller, O. J. 1977. Frequency of satellite association of human chromosomes is correlated with amount of Ag-staining of the nucleolus organizer region. Am. J. Hum. Genet 29:490-502.
  27. Pich, A., Chiusa L. and Margaria, E. 2000. Prognostic relevance of AgNORs in tumor pathology. Micron 31:133-141.
  28. Schwarzacher, H. G., Mikelsaar, A. V. and Schnedl, W. 1978. The nature of Ag-staining of nucleolus organizer regions. Electron- and light-microscopic studies on human cells in interphase, mitosis, and meiosis. Cytogenet. Cell Genet. 20:24-39.
  29. Schwarzacher, T., Mayr, B. and Schweizer, D. 1984. Heterochromatin and nucleolus organizer region behaviour at male pachytene of Sus scrofa domestica. chromosoma 91:12-19.
  30. Stefanova, V. N. 1984. Quantitative characteristics of the nucleolus organizer region of the chromosomes in the domestic pig. Tsitologiia 26:40-45.
  31. Tantravahi, U., Breg, W. R., Wertelelecki, V., Erlanger, B. F. and Miller, O. 1981. Evidence for methylation of inactive human rRNA genes in amplified regions. Hum. Genet. 56:315-320.
  32. Toga-Piquet, C., Henderson, A. S., Grillo, J. M. and Stahl, A. 1984. Localization des genes ribosomique et activite nucleolaire dans les lymphocytes du porc (Sus scrofa domestica) stimules par la phytohemagglutinine. Comptes Rendus des Seances de l'Academie des Sciences(Ⅲ) 289:383-6.
  33. Trere, D. 2000. AgNOR staining and quantification. Micron 31:127-131
  34. Zakharov, A. F., Davudov, A. Z., Benjush, V. A. and Egolina, N. A. 1982. Polymorphism of Ag-stained nucleolus organizer region in man. Human Genetics 60:334-339.
  35. Zurita, F., Jimenez, R., Burgos, M. and Diaz, del la G. R. 1998. Sequential silver staining and in situ hybridization reveal a direct association between rDNA levels and the expression of homologous nucleolar organizing regions, a hypothesis for NOR structure and function. J. Cell Sci. 111:1433-1439.
  36. 손시환, 이재익. 1998. 고분염 분석법(high-resolution banding)에 의한 한우 염색체의 표준 표지 설정. 한국축산학회지 40(5):467-484.