Skip to main content
Log in

Tools of the trade: diagnostics and research in domestic animal cytogenetics

  • Review Article
  • Published:
Journal of Applied Genetics Aims and scope Submit manuscript

Abstract

This paper describes the most common cytogenetic techniques we routinely adopt in our laboratories for producing high-resolution banding on prometaphase stage chromosomes, from synchronized or nonsynchronized blood cultures. Special emphasis is given to the FISH procedures applied to prometaphase chromosomes for mapping purposes. Each section includes historical information, basic principles for the given technique, its primary use in veterinary cytogenetics, and major limitations. Supplementary material (protocols and chemicals used) are available on our website. Even though these techniques mainly refer to the Bovidae, they can be easily extended and adapted to members of other taxa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Brunner RM, Goldammer T, Furbass R, Vanselow J, Schwerin M, 1998. Genomic organization of the bovine aromatase encoding gene and homologous pseudogene as revealed by DNA fiber FISH. Cytogenet Cell Genet 82: 37–40.

    Article  CAS  PubMed  Google Scholar 

  • CSKBB, 1994. Committee for standardized karyotype of river buffalo (Bubalus bubalis, 2n=50), (L. Iannuzzi, coordinator). Cytogenet Cell Genet 67: 102–113.

    Article  Google Scholar 

  • CSKDP, 1988. Committee for the standardized karyotype of the domestic pig (Gustavsson I, co-ordinator). Hereditas 109: 151–157.

    Google Scholar 

  • De Grouchy J, Roubin M, Passage E, 1964. Microtechnique pour l’etude des chromosomes humaines a partir d’une culture de leucocytes sanguins. Ann Génét 7: 45.

    Google Scholar 

  • Di Berardino D, Iannuzzi L, Lioi MB, 1985. The high resolution RBA-banding pattern of bovine chromosomes. Cytogenet Cell Genet 39: 136–139.

    Article  PubMed  Google Scholar 

  • Di Berardino D, Vozdova M, Kubickova S, Cernohorska H, Coppola G, Coppola GF, et al. 2004. Sexing river buffalo (Bubalus bubalis L.), sheep (Ovis aries L.), goat (Capra hircus L.) and cattle spermatozoa by double color FISH using bovine (Bos taurus L.) X- and Y-painting probes. Mol Reprod Dev 67: 108–115.

    Article  PubMed  Google Scholar 

  • Di Meo GP, Gallagher DS, Perucatti A, Wu X, Incarnato D, Mohammadi G, 2006a. Mapping of 11 genes by FISH to BTA2, BBU2q, OAR2q and CHI2, and comparison with HSA2q. Anim Genet 37: 299–300.

    Article  PubMed  Google Scholar 

  • Di Meo GP, Perucatti A, Chaves R, Adega F, De Lorenzi L, Molteni L, et al. 2006b. Cattle rob(1;29) originating from complex chromosome rearrangements as revealed by both banding and FISH-mapping techniques. Chomosome Res 14: 649–55.

    Article  CAS  Google Scholar 

  • Di Meo GP, Perucatti A, Floriot S, Hayes H, Schibler L, Rullo R, et al. 2007: An advanced sheep (Ovis aries, 2n=54) cytogenetic map and assignment of 88 new loci by fluorescent in situ hybridization and R-banding. Anim Genet 38: 233–240.

    Article  PubMed  Google Scholar 

  • Dutrillaux B, Viegas-Pequignot E, 1981: High resolution R and G banding on the same preparation. Hum Genet 57: 93–95.

    Article  CAS  PubMed  Google Scholar 

  • Evans HJ, Buckland RA, Sumner AT, 1973. Chromosome homology and heterochromatin in goat, sheep and ox studied by banding technique. Chromosoma 42: 283–402.

    Article  Google Scholar 

  • Ford CE, Pollock DL, Gustavsson I, 1980. Proceedings of the first international conference for the standardisation of banded karyotypes of domestic animals. Hereditas 92: 145–162.

    Article  CAS  PubMed  Google Scholar 

  • Gall JG, Pardue ML, 1969. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci (Wash.) 63: 378–383.

    Article  CAS  Google Scholar 

  • Gallagher DS, Basrur PK, Womack JE, 1992. Identification of an autosome to X chromosome translocation in the domestic cow. J Hered 83: 451–453.

    PubMed  Google Scholar 

  • Gallagher DS Jr, Ryan AM, Liou LS, Sastry KN, Womack JE, 1993. Somatic cell mapping of conglutinin (CGN1) to cattle syntenic group U29 and fluorescence in situ localization to chromosome 28. Mamm Genome 4: 716–719.

    Article  CAS  PubMed  Google Scholar 

  • Goldammer T, Brunner R, Weikard R, Kuehn C, Wimmers K, 2007. Generation of an improved cytogenetic and comparative map ofBos taurus chromosome BTA27. Chromosome Res 15: 203–213.

    Article  CAS  PubMed  Google Scholar 

  • Greaves IK, Rangasamy D, Ridgway P, Tremethick DJ, 2007. H2A.Z contributes to the unique 3D structure of the centromere. Proc Natl Acad Sci USA 104: 525–530.

    Article  CAS  PubMed  Google Scholar 

  • Gustavsson I, Rockborn G, 1964. Chromosome abnormality in three cases of lymphatic leukaemia in cattle. Nature 203: 990.

    Article  CAS  PubMed  Google Scholar 

  • Gustavsson I, 1969. Cytogenetics, distribution and phenotypic effects of a translocation in Swedish cattle. Hereditas 63: 68–169.

    Article  CAS  PubMed  Google Scholar 

  • Gustavsson I, Hageltorn M, Johansson C, Zech L, 1972. Identification of the pig chromosomes by the quinacrine mustard fluorescence technique. Exp Cell Res 70: 471–474

    Article  CAS  PubMed  Google Scholar 

  • Gustavsson I, 1980. Banding techniques in chromosome analysis of domestic animals. Adv Vet Sci Comp Med 24: 245–289.

    CAS  PubMed  Google Scholar 

  • Gustavsson I, Switonski M, Larsson K, Ploen L, Hojer K, 1988. Chromosome banding studies and synaptonemal complex analyses of four reciprocal translocations in the domestic pig. Hereditas 109: 169–184.

    Article  CAS  PubMed  Google Scholar 

  • Hansen KM, 1971. Identification of bovine chromosomes by the quinacrine mustard fluorescence technique. Hereditas 69: 295.

    Google Scholar 

  • Hansen KM, 1973. Heterochromatin (C-bands) in bovine chromosomes. Hereditas 73: 65–70.

    Article  CAS  PubMed  Google Scholar 

  • Hayes H, Petit E, Bouniol C, Popescu P, 1993. Localization of the alpha-S2-casein gene (CASAS2) to the homologous cattle, sheep and goat chromosomes 4 by in situ hybridization. Cytogenet Cell Genet 64: 281–285.

    Article  CAS  PubMed  Google Scholar 

  • Hayes H, Di Meo GP, Gautier M, Laurent P, Eggen A, Iannuzzi L, 2000. Localization by FISH of the 31 Texas nomenclature type I markers to both Q- and R-banded bovine chromosomes. Cytogenet Cell Genet 90: 315–320.

    Article  CAS  PubMed  Google Scholar 

  • Hayes H, Rogel-Gaillard C, Zijlstra C, De Haan NA, Urien C, Bourgeaux N, 2002. Establishment of an R-banded rabbit karyotype nomenclature by FISH localization of 23 chromosome-specific genes on both G- and R-banded chromosomes. Cytogenet Genome Res 98: 199–205.

    Article  CAS  PubMed  Google Scholar 

  • Henderson LM, Bruere AN, 1977. Association of nucleolus organizer regions in domestic sheep (Ovis aries) shown by silver staining. Cytogenet Cell Genet 19: 326–334.

    Article  CAS  PubMed  Google Scholar 

  • Hsu TC, Pomerat CM, 1953. Mammalian chromosomes in vitro II. A method for spreading the chromosomes of cells in tissue culture. J Hered 44: 23–29.

    Google Scholar 

  • Iannuzzi L, 1990. An improved characterization of cattle chromosomes by means of high resolution G-and R-band comparison. J Hered 81: 80–83.

    CAS  PubMed  Google Scholar 

  • Iannuzzi L, Di Meo GP, Perucatti A, Ferrara L, 1990. The high resolution G- and R-banding patterns in chromosomes of river buffalo (Bubalus bubalis L.). Hereditas 112: 209–215.

    Article  CAS  PubMed  Google Scholar 

  • Iannuzzi L, Di Meo GP, Gallagher DS, Ryan AM, Ferrara L, Womack JE, 1993. Chromosomal localization of omega and trophoblast interferon genes in goat and sheep by fluorescent in situ hybridization. J Hered 84: 301–304.

    CAS  PubMed  Google Scholar 

  • Iannuzzi L, Di Meo GP, Perucatti A, Incarnato D, 1999. Comparison of the human with the sheep genomes by use of human chromosome-specific painting probes. Mamm Genome 10: 719–723.

    Article  CAS  PubMed  Google Scholar 

  • Iannuzzi L, Di Meo GP, Perucatti A, Rullo R, Incarnato D, Longevi M, et al. 2003. Comparative FISH-mapping of the survival of motor neuron gene (SMN) in domestic bovids. Cytogenet Genome Res 102: 39–41.

    Article  CAS  PubMed  Google Scholar 

  • Iannuzzi L, Di Meo GP, Perucatti A, Incarnato D, Di Palo R, Zicarelli L, 2004. Reproductive disturbances and sex chromosome abnormalities in river buffaloes. Vet Rec 154: 823–824.

    Article  CAS  PubMed  Google Scholar 

  • ISCNDA, 1990. International System for Cytogenetic Nomenclature of Domestic Animals (Di Berardino D, Hayes H, Fries R, Long S, eds). Cytogenet Cell Genet 53: 65–79.

  • ISCNH, 1997. International System for Cytogenetic Nomenclature of the Domestic Horse, Bowling AT, Breen M, Chowdhary BP, Hirota K, Lear T, Millon LV, Ponce de Leon FA, Raudsepp T, Stranzinger G. (Committee). Chromosome Res 5: 433–443.

    Article  Google Scholar 

  • ISCNDB, 2001. International System for Chromosome Nomenclature of Domestic Bovids. Di Berardino D, Di Meo GP, Gallagher DS, Hayes H, Iannuzzi L (co-ordinator) eds. Cytogenet Cell Genet 92: 283–299.

  • Landegent JE, Jansen IWN, Dirks RW, Baas F, van der Ploeg M, 1987. Use of whole cosmid cloned genetic sequences for chromosomal localization by non-radioactive in situ hybridization. Hum Genet 77: 366–370.

    Article  CAS  PubMed  Google Scholar 

  • Langer PR, Waldrop AA, Ward DC, 1981. Enzymatic synthesis of biotin-labelled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78: 6633–6637.

    Article  CAS  PubMed  Google Scholar 

  • Makino S, Sasaki MS, Sofuni T, Ishikawa T, 1962. Chromosome condition of an intersex swine. Proc Jpn Acad 38: 686–689.

    Google Scholar 

  • McConnell J, Feccheimer NS, Gilmore LO, 1963. Somatic chromosomes of the domestic pig. J Anim Sci 22: 374–379.

    Google Scholar 

  • Molteni L, Perucatti A, Iannuzzi A, Di Meo GP, De Lorenzi L, De Giovanni A, 2007. A new case of reciprocal translocation in a young bull: rcp(11;21)(q28;q12). Cytogenet Genome Res 116: 80–84.

    Article  CAS  PubMed  Google Scholar 

  • Moorhead PS, Nowell PC, Mellman WJ, Battips DM, Hundgerford DA, 1960. Chromosome preparations of leucocytes cultured from human peripheral blood. Exp Cell Res 20: 613–616.

    Article  CAS  PubMed  Google Scholar 

  • Pinkel D, Straume T, Gray JW, 1986. Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83: 2934–2938.

    Article  CAS  PubMed  Google Scholar 

  • Popescu CP, 1975. Essai d’identification des chromosomes bovins (Bos taurus) à l’aide du marquage au 5-bromodeoxyuridine (BrdU). I « 2e Colloque Européen de Cytogénétique des animaux domestiques, Giessen, 29–30 September: 59–64.

  • Poulsen BS, Shibasaki Y, Ronne M, 1988. The high resolution R-banded karyotype ofOryctolagus cuniculus L. Hereditas 109: 57–60.

    Article  CAS  PubMed  Google Scholar 

  • Romagnano A, Richer CL, 1985. High resolution R-bands produced in equine chromosomes after incorporation of bromodeoxyuridine. J Hered 76: 377–378.

    CAS  PubMed  Google Scholar 

  • Ronne M, 1984. Fluorouracil synchronization of human lymphocyte cultures. Induction of high resolution R-banding by simultaneous exposure to 5-bromodeoxyuridine/Hoechst 33258 in vitro. Hereditas 101: 205–208

    Article  CAS  PubMed  Google Scholar 

  • Ronne M, Andersen O, Hansen SO, 1984. Methotrexate-leucovorin synchronization of human lymphocyte cultures. Induction of high resolution R- and G-banding. Anticancer Res 4: 357–360

    CAS  PubMed  Google Scholar 

  • Ronne M, 1990. The high resolution R-banded karyotype ofSus scrofa domestica L: In vivo 4: 181–184.

    CAS  PubMed  Google Scholar 

  • Ruechert RR, Mueller GC, 1960. Studies on unbalanced growth in tissue cultures. I. Induction and consequences of thymidine deficiency. Cancer Res 20: 1584–1591.

    Google Scholar 

  • Schibler L, Vaiman D, Oustry A, Giraud-Delville C, Cribiu EP, 1998. Comparative gene mapping: a fine-scale survey of chromosome rearrangements between ruminants and humans. Genome Res 8: 901–915.

    CAS  PubMed  Google Scholar 

  • Stone E, 1963. A chromosome analysis of the domestic pig (Sus scrofa) utilizing a peripheral blood culture technique. Can J Genet Cytol 5: 38–42.

    CAS  PubMed  Google Scholar 

  • Switonski M, Reimann N, Bosma AA, Long S, Bartnitzke S, Pienkowska A, 1996. Report on the progress of standardization of the G-banded canine (Canis familiaris) karyotype. Chromosome Res 4: 306–309.

    Article  CAS  PubMed  Google Scholar 

  • Texas Nomenclature, 1996. Standardization of cattle karyotype nomenclature: report of the committee for the standardization of the cattle karyotype (Popescu CP, Long S, Riggs P, Womack J, Schmutz S, Fries R, Gallagher D, eds). Cytogenet Cell Genet 74: 259–261.

  • Viegas-Pequignot E, Dutrillaux B, 1978. Une method simple pour obtenir des prophases et des prometaphases. Ann Génét 21: 122–125.

    Google Scholar 

  • Webber LM, Garson MO, 1983. Fluorodeoxyuridine synchronization of bone marrow cultures. Cancer Genet Cytogen 8: 123–132.

    Article  CAS  Google Scholar 

  • Yunis JJ, 1976. High resolution of human chromosomes. Science 191: 1268–1270.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Dino Di Berardino.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Iannuzzi, L., Di Berardino, D. Tools of the trade: diagnostics and research in domestic animal cytogenetics. J Appl Genet 49, 357–366 (2008). https://doi.org/10.1007/BF03195634

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03195634

Keywords

Navigation