, Volume 133, Issue 3, pp 261–267 | Cite as

Variation in size and location of the Ag-NOR in the Atlantic halibut (Hippoglossus hippoglossus)

  • K. Ocalewicz
  • D. J. Penman
  • I. Babiak


The distribution of differentially stained chromatin was studied in the Atlantic halibut (Hippoglossus hippoglossus) chromosomes (2n = 48). Four pairs of homologous chromosomes were identified using a combination of traditional cytogenetic staining techniques (Giemsa/DAPI/CMA3/Ag-NO3). Chromosome 1 showed a length polymorphism (1S-short, 1L-long isoforms of the chromosome 1) which was related to the variation of the size of the Ag-NORs. In one specimen the Ag-NOR was translocated from chromosome 1 into the telomeric region on the q-arm of the chromosome 2 forming a derivative chromosome der(2)t(1S;2)(q?;q?). Four Ag-NOR genotypes have been shown: 1S1S, 1S1L, 1L1L and 1S der(2)t(1S;2)(q?;q?). The chromosome rearrangements did not leave any interstitially located telomeric sequences and the telomeres were confined to the ends of the chromosomes. A single chromosomal location of 5S rDNA clusters was found using the PRINS technique. In the extended metaphase spreads two adjacent clusters of 5S rDNA could be seen on one chromosome while condensed chromatin gave a single hybridization signal. Double 5S rDNA signals on the same chromosome arm suggested paracentric inversion of the minor rDNA site. 5S rDNA clusters were not co-localized with Ag-NORs. Although female and male karyotypes were compared no sex related cytogenetic markers were found.


Chromosomes DAPI NORs Polymorphism Atlantic halibut Sex 



This work was supported by research project no. 165272/S40 financed by the Research Council of Norway.


  1. Amemiya ChT, Gold JR (1986) Chromomycin A3 stains Nucleolus Organizer Regions of fish chromosomes. Copeia 1:226–231CrossRefGoogle Scholar
  2. Appels R, Gerlach WL, Dennis ES et al. (1980) Molecular and hromosomal organization of DNA sequencing coding for the ribosomal RNAs in cereals. Chromosoma 78:293–311CrossRefGoogle Scholar
  3. Azevedo MFC, Oliveira C, Pardo BG et al. (2007) Cytogenetic characterization of six species of flatfishes with comments to karyotype differentiation patterns in Pleuronectiformes (Teleostei). J Fish Biol 70:1–15CrossRefGoogle Scholar
  4. Barker CJ (1972) A new method for the display of chromosome plaice, Pleuronectes platessa, and other marine fishes. Copeia 2:365–368CrossRefGoogle Scholar
  5. Bjørnsson B (1995) The growth pattern and sexual maturation of Atlantic halibut (Hippoglossus hippoglossus L.) rerared in large tanks for 3 years. Aquaculture 138:281–290CrossRefGoogle Scholar
  6. Brown NP, Bromage NR, Penman DJ (1997) The karyotype of Atlantic halibut, Hippoglossus hippoglossus (Linnaeus). Aquac Res 28:489–491CrossRefGoogle Scholar
  7. Cross I, Merlo A, Manchado M et al. (2006) Cytogenetic characterization of the sole Solea senegalensis (Teleostei: Pleuronectiformes: Soleidae): Ag-NOR, (GATA)n, (TTAGGG)n and ribosomal genes by one-color and two-color FISH. Genetica 128:253–259PubMedCrossRefGoogle Scholar
  8. De Lucchini S, Andronico F, Nardi I (1997) Molecular structure of the rDNA intergenic spacer (IGS) in Triturus: implications for the hypervariability of rDNA loci. Chromosoma 106:315–326PubMedCrossRefGoogle Scholar
  9. Fujiwara A, Fujiwara M, Nishida, C et al. (2007) Characterization of Japanese flounder by chromosome bandings and fluorescence in situ hybridization with DNA markers. Genetica: DOI  10.1007/s10709-006-9136-z
  10. Garrido-Ramos MA, Jamilena M, Lozano R et al. (1995) Cytogenetic analysis of gilthead seabram Sparus auratus (Pisces, Perciformes), a deletion affecting the NOR in a hatchery stock. Cytogenet Cell Genet 68:3–7PubMedCrossRefGoogle Scholar
  11. Howell WM, Black DA (1980) Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia 36:1014–1015PubMedCrossRefGoogle Scholar
  12. ISCN (1995). International System for human cytogenetic nomenclature. Mitelman F, editor. Basel: S. Karger, 1995Google Scholar
  13. Jankun M, Ocalewicz K, Pardo BG et al. (2003) Chromosomal characteristics of rDNA in European grayling Thymallus thymallus (Salmonidae). Genetica 119:219–224PubMedCrossRefGoogle Scholar
  14. Kavalco KF, Pazza R, Bertollo LAC et al. (2004) Gene mapping of 5S rDNA sites in eight fish species from the Paraiba do Sl river basin, Brazil. Cytogenet. Genome Res. 106:107–110Google Scholar
  15. Koch J, Kolvraa S, Petersen K et al. (1989) Oligonucleotide-priming methods for chromosome specific labeling of alpha satellite DNA in situ. Chromosoma 98:259–265PubMedCrossRefGoogle Scholar
  16. Kress H, Bechler K, Swida U, Maletz S (2001). Evolution of 5S rRNA gene families in Drosophila. Chromosome Res 9:403–15PubMedCrossRefGoogle Scholar
  17. Lee SH, Do GS, Seo BB (1999) Chromosomal localization of 5S rRNA gene loci and the implications for relationships within the Allium complex. Chromosome Res 7:89–93PubMedCrossRefGoogle Scholar
  18. Le François NR, Lemieux H, Blier PU (2002) Biological and technical evaluation of the potential of marine and anadromous fish species for cold-water mariculture. Aquac Res 33:95–108CrossRefGoogle Scholar
  19. LeGrande WH (1975) Karyology of six species of Louisiana flatfishes (Pleuronectiformes: Osteichthyes). Copeia 1975:516–522CrossRefGoogle Scholar
  20. Manchado M, Zuasti E, Cross I et al. (2005) Molecular characterization and chromosomal mapping of the 5S rRNA gene in Solea senegalensis: a new limkage to the U1, U2 and U5 small nuclear RNA genes. Genome 49(1):79–86CrossRefGoogle Scholar
  21. Martins G, Galetti PM (1999) Chromosomal localization of 5S rRNA genes in Leporinus fish (Anastomidae, Characiformes). Chromosome Res 7:363–367PubMedCrossRefGoogle Scholar
  22. Mukai Y, Endo TR, Gill BS (1990) Physical mapping of the 5S rRNA multigene family in common wheat. Heredity 81:290–295Google Scholar
  23. Ocalewicz K, Babiak I (2003) Primed in situ labeling (PRINS) detection of 5S rDNA sequences proves absence of X chromosome in supermale androgenetic rainbow trout. J Fish Biol 62:1462–1466CrossRefGoogle Scholar
  24. Ocalewicz K, Hliwa P, Krol J et al. (2007) Karyotype and chromosomal characteristics of Ag-NOR sites and 5S rDNA in European smelt (Osmerus eperlanus L.). Genetica 131:29–35PubMedCrossRefGoogle Scholar
  25. Pardo MBG, Bouza C, Castro J et al. (2001) Localization of ribosomal genes in Pleuronectiformes using Ag-, CMA3-banding and in situ hybridization. Heredity 86:531–536PubMedCrossRefGoogle Scholar
  26. Park IS, Nam YK, Douglas SE, Johnson SC et al. (2003) Genetic characterization, morphometrics and gonad development of induced interspecific hybrids between yellowtail flounder, Pleuronectes ferrugineus (Storer) and winter flounder, Pleuronectes americanus (Walbaum). Aquac Res 34:389–396CrossRefGoogle Scholar
  27. Ráb P, Roth P (1988) Cold-blooded vertebrates. In: Balicek P, Forejt J, Rubes J (Eds) Methods of chromosome analysis, Brno, Cytogeneticka sekce Ceskoslovenske biologicke spolecnosti pri CSAV, Czech Republic, pp 115–124Google Scholar
  28. Sola L, Cataudella S, Capanna E (1981) New developments in vertebrate cytotaxonomy. III. Karyology of bony fishes: a review. Genetica 54:285–328CrossRefGoogle Scholar
  29. Sola L, Rossi AR, Iaselli V et al. (1992) Cytogenetics of bisexual/unisexual species of Poecilia. II. Analysis of heterochromatin and nucleolar organizer regions in Poecilia mexicana mexicana by C-banding and DAPI, quinacrine, chromomycin A3, and silver staining. Cytogenet Cell Genet 60:229–235PubMedCrossRefGoogle Scholar
  30. Suzuki H, Sakurai S, Matsuda Y (1996) Rat rDNA spacer sequences and chromosomal assignment of the genes to the extreme terminal region of chromosome 19, Cytogenet Cell Genet 72:1–4PubMedCrossRefGoogle Scholar
  31. Vasiliev VP (1978) Karyotypes of 5 species of fishes (Pisces) from the Black Sea. Tsitologiya 20:1092–1094Google Scholar
  32. Vitturi R, Catalano R, Colombera D (1993) Chromosome analysis of Bothus podas (Pisces, Pleuronectiformes) from the Mediterranean Sea. J Fish Biol 43:221–227Google Scholar
  33. Winkler FM, García-Melys D, Palma-Rojas C (2004) Karyotypes of three South East Pacific flounder species of the family Paralichthyidae. Aquac Res 35:1295–1298CrossRefGoogle Scholar
  34. Zurita F, Jimenez R, Burgos M et al. (1998) Sequential silver staining and in situ hybridization reveal a direct association between rDNA levels and the expression of homologous nuclear organizing regions: a hypothesis for NOR structure and function. J Cell Sci 111:1433–1439PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Department of IchthyologyUniversity of Warmia and Mazury in OlsztynOlsztynPoland
  2. 2.Department of Fisheries and Natural SciencesBodø University CollegeBodoNorway
  3. 3.Institute of AquacultureUniversity of StirlingStirlingUK

Personalised recommendations