, Volume 103, Issue 1, pp 31–39 | Cite as

Sex chromosome polymorphism and heterogametic males revealed by two cloned DNA probes in the ZW/ZZ fish Leporinus elongatus

  • Ichiro Nakayama
  • Fausto Foresti
  • Rita Tewari
  • Manfred Schartl
  • Daniel Chourrout
Original Articles


In order to study the divergence of teleost sex chromosomes, subtractive cloning was carried out between genomic DNA of males and females of the rainbow trout (XX/XY) and of Leporinus elongatus (ZW/ZZ). Inserts cloned in a plasmid vector were individually tested on Southern blots of DNA of males and females for sex specificity. No sex-specific insert was obtained from trout, but two out of ten inserts cloned from L. elongatus showed sex-specific patterns in this species: one corresponds to a sequence present on both Z and W chromosomes, while the other is W specific. Sequences of these two inserts show neither clear homology with other known sequences, nor an open reading frame. They cross-hybridize with the genomic DNA of Leporinus friderici, but without sex-specific patterns. Twenty-four L. elongatus adults were sexed by gonadal observation, chromosomed examination and Southern hybridization with one or the other insert. Ten males and 11 females had chromosomes and hybridization patterns typical of their sex. One ZW female was recognized as a male with the W-specific probe. This was also the case for two unusual ZW males, one having a male hybridization pattern with the other probe. These three atypical individuals may result from single genetic exchanges between four regions of the Z and the W, giving rise to three atypical W chromosomes. Finding males with such atypical heterochromosomes in a female heterogametic species may indicate that a gradual transition occurs between the heterogametic systems.


Rainbow Trout Southern Blot Plasmid Vector Southern Hybridization Gradual Transition 
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.


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  1. Aida T (1921) On the inheritance of color in a fresh water fish, Aplocheilus latipes, with special reference to sex-linked inheritance. Genetics 6:554–573Google Scholar
  2. Beçak W (1983) Evolution and differentiation of sex chromosomes in lower vertebrates. In: Müller U, Franke WW (eds) Mechanism of gonadal differentiation in vertebrates. Differentiation [Suppl] 23:S3–12Google Scholar
  3. Charlesworth B (1991) The evolution of sex chromosomes. Science 251:1030–1033Google Scholar
  4. Chourrout D, Quillet E (1982) Induced gynogenesis in the rainbow trout: sex and survival of progenies. Production of all-triploid populations. Theor Appl Genet 63:201–205Google Scholar
  5. Clemens HP, Inslee T (1968) The production of unisexual broods of Tilapia mossambica sex-reversed with methyltestosterone. Trans Am Fish Soc 97:18–21Google Scholar
  6. Cotinot C, Kirszenbaum M, Leonard M, Gianquinto L, Vaiman M (1991) Isolation of bovine Y-derived sequence: potential use in embryo sexing. Genomics 10:646–653Google Scholar
  7. de la Chapelle A (1981) Etiology of maleness in XX men. Hum Genet 58:105–116Google Scholar
  8. Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395Google Scholar
  9. Devlin RH, McNeil BK, Groves TDD, Donaldson EM (1991) Isolation of a Y-chromosomal DNA probe capable of determining genetic sex in chinook salmon (Oncorhynchus tshawytscha). Can J Fish Aquat Sci 48:1606–1612Google Scholar
  10. Disteche CM, Casanova M, Saal H, Friedman C, Sybert V, Graham J, Thuline H, Page DC, Fellous M (1986) Small deletions of the short arm of the Y chromosome in 46, XY females. Proc Natl Acad Sci USA 83:7841–7844Google Scholar
  11. Ferreiro C, Medrano JF, Gall GAE (1989) Genome analysis of rainbow trout and sturgeon with restriction enzymes and hybridization with a ZFY gene derived probe to identify sex. Aquaculture 81:245–251Google Scholar
  12. Foerster W, Anders F (1977) Zytogenetischer Vergleich der Karyotypen verschidener Rassen und Arten lebendgebärender Zahnkarpfen der Gattung Xiphophorus. Zool Anz 198:167–177Google Scholar
  13. Fredga K, Gropp A, Winking H, Frank F (1976) Fertile XX and XY type females in wood lemming, Myopus schisticolor. Nature 261:225–227Google Scholar
  14. Galetti PM Jr, Foresti F (1986) Evolution of the ZZ/ZW system in Leporinus (Pisces, Anostomidae). Cytogenet Cell Genet 43:43–46Google Scholar
  15. Galetti PM Jr, Foresti F, Bertollo LAC, Moreira Filho O (1981) Heteromorphic sex chromosomes in three species of the genus Leporinus (Pisces, Anostomidae). Cytogenet Cell Genet 29:138–142Google Scholar
  16. Galetti PM Jr, Foresti F, Bertollo LAC, Moreira Filho O (1984) Characterization of eight species of Anostomidae (Cypriniformes) fish on the basis of the nucleolar organizing region. Caryologia 37:401–406Google Scholar
  17. Gordon M (1947)_Genetics of Platypoecilius maculatus. IV. The sex-determining mechanism in two wild populations of the Mexican platy fish. Genetics 32:8–17Google Scholar
  18. Guerrero RD (1975) Use of androgens for the production of allmale Tilapia aurea (Steindachner). Trans Am Fish Soc 2:342–348Google Scholar
  19. Haaf T, Schmid M (1984) An early stage of ZW/ZZ sex chromosome differentiation in Poecilia sphenops var. melanistica (Poeciliidae, Cyprinodontiformes). Chromosoma 89:37–41Google Scholar
  20. Hickling CF (1960) The Malacca tilapia hybrids. J Genet 57:1–10Google Scholar
  21. Hunter GA, Donaldson EM (1983) Hormonal sex control and its application to fish culture. In: Hoar WS, Randall DJ, Donaldson EM (eds) Fish physiology, vol 9B. Academic Press, New York, pp 223–303Google Scholar
  22. Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Improved technique utilizing non fat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Genet Anal Tech 1:3–8Google Scholar
  23. Kallman KD (1984) A new look at sex determination in poeciliid fishes. In: Turner BJ (ed) Evolutionary genetics of fishes. Plenum Press, New York, pp 95–171Google Scholar
  24. Kessler C, Höltke HJ, Seibl R, Burg J, Mühlegger K (1990) Nonradioactive labelling and detection of nucleic acids. I. A novel DNA labelling and detection system based on digoxigenin: antidigoxigenin elisa principle (digoxigenin system). Biol Chem Hoppe-Seyler 371:917–927Google Scholar
  25. Lamar EE, Palmer E (1984) Y-encoded, species specific DNA in mice: evidence that the Y-chromosome exists in two polymorphic forms in inbred strains. Cell 37:171–177Google Scholar
  26. Lloyd MA, Fields MJ, Thorgaard GH (1989) Bkm minisatellite sequences are not sex associated but reveal DNA fingerprint polymorphism in rainbow trout. Genome 32:865–868Google Scholar
  27. Nagy A, Rajki K, Horvath L, Csanyi V (1978) Investigation on carp, Cyprinus carpio, L. gynogenesis. J Fish Biol 13:215–224Google Scholar
  28. Nanda I, Feichtinger W, Schmid M, Schröder JH, Zischler H, Epplen JT (1990) Simple repetitive sequences are associated with differentiation of the sex chromosomes in the sex chromosomes in the guppy fish. J Mol Evol 30:456–462Google Scholar
  29. Nanda I, Schartl M, Feichtinger W, Epplen JT, Schmid M (1992) Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences. Chromosoma 101:301–310Google Scholar
  30. Price DJ (1984) Genetics of sex determination in fishe — a brief review. In: Fish reproduction. Academic Press, New York, pp 77–89Google Scholar
  31. Rouyer F, Simmler MC, Page DC, Weissenbach J (1987) A sex chromosome rearrangement in a human XX male caused by Alu-Alu recombination. Cell 51:417–425Google Scholar
  32. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  33. Sanger F, Nicklen S, Coulson AP (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  34. Sibl R, Höltke HJ, Rüger R, Meindl A, Zachau HG, Rasshoffer R, Roggendorf M, Wolf H, Arnold N, Weinberg J, Kessler C (1990) Non radioactive labelling and detection of nucleic acids. III. Applications of the digoxigenin system. Biol Chem Hoope-Seyler 371:939–951Google Scholar
  35. Smith CL (1975) The evolution of hermaphroditism in fishes. In: Reinboth R (ed) Intersexuality in the animal kingdom. Springer, Berlin Heidelberg New York, pp 295–310Google Scholar
  36. Sola L, Cataudella S, Capanna E (1981) New developments in vertebrate cytotaxonomy III. Karyology of bony fishes: a review. Genetica 54:285–328Google Scholar
  37. Southern E (1975) Detection of specific sequences among DNA fragments separated by agarose gels. J Mol Biol 98:503–517Google Scholar
  38. Thorgaard GH (1977) Heteromorphic sex chromosomes in male rainbow trout. Science 196:900–902Google Scholar
  39. Thorgaard GH (1983) Chromosomal differences among rainbow trout, Salmo gairdneri populations. Copeia 1983:650–662Google Scholar
  40. Yamamoto T (1953) Artificially induced sex reversal in genotypic males of the medaka (Oryzias latipes). J Exp Zool 123:571–594Google Scholar
  41. Yamamoto T (1955) Progeny of artificially induced sex reversal of male genotype (XY) in the medaka (Oryzias latipes) with special reference to YY male. Genetics 40:406–419Google Scholar
  42. Zhang FR, Heilig R, Thomas G, Aurias A (1990) One step efficient and specific non radioactive non fluorescent method for in situ hybridisation of banded chromosomes. Chromosoma 99:436–439Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Ichiro Nakayama
    • 1
  • Fausto Foresti
    • 2
  • Rita Tewari
    • 1
  • Manfred Schartl
    • 3
  • Daniel Chourrout
    • 1
  1. 1.Laboratoire de Génétique des PoissonsInstitut National de la Recherche AgronomiqueJouy en JosasFrance
  2. 2.Departamento de Morfologia, CEPUmversidade Estadual Paulista “Julio de Mesquita Filho”BotucatuBrazil
  3. 3.Lehrstuhl Physiologische Chemie ITheodor-Boveri-Institut für Biowissenschaften der Universität WürzburgWürzburgGermany

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