, 120:455 | Cite as

Strong conservation of the bird Z chromosome in reptilian genomes is revealed by comparative painting despite 275 million years divergence

  • Martina Pokorná
  • Massimo Giovannotti
  • Lukáš KratochvílEmail author
  • Fumio Kasai
  • Vladimir A. Trifonov
  • Patricia C. M. O’Brien
  • Vincenzo Caputo
  • Ettore Olmo
  • Malcolm A. Ferguson-Smith
  • Willem RensEmail author
Research Article


The divergence of lineages leading to extant squamate reptiles (lizards, snakes, and amphisbaenians) and birds occurred about 275 million years ago. Birds, unlike squamates, have karyotypes that are typified by the presence of a number of very small chromosomes. Hence, a number of chromosome rearrangements might be expected between bird and squamate genomes. We used chromosome-specific DNA from flow-sorted chicken (Gallus gallus) Z sex chromosomes as a probe in cross-species hybridization to metaphase spreads of 28 species from 17 families representing most main squamate lineages and single species of crocodiles and turtles. In all but one case, the Z chromosome was conserved intact despite very ancient divergence of sauropsid lineages. Furthermore, the probe painted an autosomal region in seven species from our sample with characterized sex chromosomes, and this provides evidence against an ancestral avian-like system of sex determination in Squamata. The avian Z chromosome synteny is, therefore, conserved albeit it is not a sex chromosome in these squamate species.


Chromosome Painting Acrocentric Chromosome Metacentric Chromosome Squamate Reptile Agamid Lizard 
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.



We are grateful to Frances L. Lovell, Margaret Wallduck, Paola Nisi Cerioni, and Šárka Pelikánová for assisting in cell culture or FISH experiments and two anonymous reviewers for valuable comments. Petr Ráb and Marie Rábová provided continuous support and encouragement. The studies were supported by grants GAČR 506/10/0718, GAUK 9942/2009 and MŠMT 0021620828. The work of MP in Cambridge was supported by the ThermAdapt research network programme of the European Science Foundation ( VAT was supported by the Royal Society and RFBR grants. The paper represents the third part of the series “Evolution of sex determining systems in lizards” by MP and LK.


  1. Andrews R (2005) Incubation temperature and sex ratio of the veiled chameleon (Chamaeleo calyptratus). J Herpetol 39:515–518CrossRefGoogle Scholar
  2. Bellott DW, Skaletsky H, Pyntikova T, Mardis ER, Graves T, Kremitzki C, Brown LG, Rozen S, Warren WC, Wilson RK, Page DC (2010) Convergent evolution of chicken Z and human X chromosomes by expansion and gene acquisition. Nature 466:612–616PubMedCrossRefGoogle Scholar
  3. Brunner B, Hornung U, Shan Z, Nanda I, Kondo M, Zend-Ajusch E, Haaf T, Ropers HH, Shima A, Schmid M, Kalscheuer VM, Schartl M (2001) Genomic organization and expression of the doublesex-related gene cluster in vertebrates and detection of putative regulatory regions for DMRT1. Genomics 77:8–17PubMedCrossRefGoogle Scholar
  4. Caputo V, Odierna G, Aprea G (1994) A chromosomal study of Eumeces and Scincus, primitive members of the Scincidae (Reptilia, Squamata). Boll Zool 61:155–162Google Scholar
  5. Chowdhary BP, Raudsepp T (2000) HSA4 and GGA4: remarkable conservation despite 300-Myr divergence. Genomics 64:102–105PubMedCrossRefGoogle Scholar
  6. Ellegren H (2010) Evolutionary stasis: the stable chromosomes of birds. Trends Ecol Evol 25:283–291PubMedCrossRefGoogle Scholar
  7. Ezaz T, Quinn AE, Miura I, Sarre D, Georges A, Graves JAM (2005) The dragon lizard Pogona vitticeps has ZZ/ZW micro-sex chromosomes. Chromosome Res 13:763–776PubMedCrossRefGoogle Scholar
  8. Ezaz T, Quinn AE, Sarre SD, O’Meally D, Georges A, Graves JAM (2009a) Molecular marker suggests rapid changes of sex-determining mechanisms in Australian dragon lizards. Chromosome Res 17:91–98PubMedCrossRefGoogle Scholar
  9. Ezaz T, Moritz B, Waters PD, Graves JAM, Georges A, Sarre SD (2009b) The ZW sex microchromosomes of an Australian dragon lizard share no homology with those of other reptiles or birds. Chromosome Res 17:965–973PubMedCrossRefGoogle Scholar
  10. Ezaz T, Sarre SD, O’Meally D, Graves JAM, Georges A (2010) Sex chromosome evolution in lizards: independent origins and rapid transitions. Cytogenet Genome Res 127:249–260CrossRefGoogle Scholar
  11. Ferguson-Smith MA, Trifonov V (2007) Mammalian karyotype evolution. Nat Rev Genet 8:950–962PubMedCrossRefGoogle Scholar
  12. Fridolfsson AK, Cheng H, Copeland NG, Jenkins NA, Liu HC, Raudsepp T, Woodage T, Chowdhary B, Halverson J, Ellegren H (1998) Evolution of the avian sex chromosomes from an ancestral pair of autosomes. Proc Natl Acad Sci USA 95:8147–8152PubMedCrossRefGoogle Scholar
  13. Gamble T, Bauer AM, Greenbaum E, Jackman TR (2008) Out of the blue: a novel, trans-Atlantic clade of gecko lizards (Gekkota, Squamata). Zool Scri 37:355–366CrossRefGoogle Scholar
  14. Giovannotti M, Caputo V, O’Brien PCM, Lovell FL, Trifonov V, Nisi Cerioni P, Olmo E, Ferguson-Smith MA, Rens W (2009) Skinks (Reptilia: Scincidae) have highly conserved karyotypes as revealed by chromosome painting. Cytogenet Genome Res 127:224–231PubMedCrossRefGoogle Scholar
  15. Glas R, Graves JAM, Toder R, Ferguson-Smith MA, O’Brien PCM (1999) Cross-species chromosome painting between human and marsupial directly demonstrates the ancient region of the mammalian X. Mamm Genome 10:1115–1116PubMedCrossRefGoogle Scholar
  16. Gorman GC, Gress F (1970) Sex chromosomes of a pygopodid lizard, Lialis burtonis. Experientia 26:206–207PubMedCrossRefGoogle Scholar
  17. Graves JAM (2009) Weird animal genomes and the evolution of vertebrate sex and sex chromosomes. Annu Rev Genet 42:565–586CrossRefGoogle Scholar
  18. Graves JAM, Shetty S (2001) Sex from W to Z: evolution of vertebrate sex chromosomes and sex determining genes. J Exp Zool 290:449–462CrossRefGoogle Scholar
  19. Itoh Y, Kampf K, Arnold AP (2006) Comparison of the chicken and zebra finch Z chromosomes shows evolutionary rearrangements. Chromosome Res 14:805–815PubMedCrossRefGoogle Scholar
  20. Ivanov VG, Bogdanov OP, Anislmova EY, Fedorova TA (1973) Studies of the karyotypes of three lizard species (Sauria, Scincidae, Lacertidae). Tsitologiya 15:1291–1296Google Scholar
  21. Janes DE, Organ CL, Fujita MK, Shedlock AM, Edwards SV (2010) Genome evolution in Reptilia, the sister group of mammals. Annu Rev Genomics Hum Genet 11:239–264PubMedCrossRefGoogle Scholar
  22. Kasai F, O’Brien PTM, Martin S, Ferguson-Smith MA (2003) Identification of the homologue of chicken Z chromosome in turtle and crocodile. Ann Génét 46:89Google Scholar
  23. Kawai A, Nishida-Umehara C, Ishijima J, Tsuda Y, Ota H, Matsuda Y (2007) Different origins of bird and reptile sex chromosomes inferred from comparative mapping of chicken Z-linked genes. Cytogenet Genome Res 117:92–102PubMedCrossRefGoogle Scholar
  24. Kawai A, Ishijima J, Nishida C, Kosaka A, Ota H, Kohno SI, Matsuda Y (2009) The ZW sex chromosomes of Gekko hokouensis (Gekkonidae, Squamata) represent highly conserved homology with those of avian species. Chromosoma 118:43–51PubMedCrossRefGoogle Scholar
  25. Kejnovský E, Leitch I, Leitch A (2009) Contrasting evolutionary dynamics between angiosperm and mammalian genomes. Trends Ecol Evol 24:572–582PubMedCrossRefGoogle Scholar
  26. King M, Mengden GA, King D (1982) A pericentric-inversion polymorphism and a ZZ/ZW sex-chromosome system in Varanus acanthurus Boulenger analyzed by G- and C-banding and Ag staining. Genetica 58:39–45CrossRefGoogle Scholar
  27. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) ClustalW and ClustalX version 2.0. Bioinformatics 23:2947–2948PubMedCrossRefGoogle Scholar
  28. Li Y, Zhang LQ, Zhang DX, Zhang XQ, Lu XM (2010) Faster evolution of Z-linked duplicate genes in chicken. J Genet Genomics 37:695–702PubMedCrossRefGoogle Scholar
  29. Matsubara K, Tarui H, Toriba M, Yamada K, Nishida-Umehara Ch, Agata K, Matsuda Y (2006) Evidence for different origin of sex chromosomes in snakes, birds, and mammals and stepwise differentiation of snake sex chromosomes. Proc Natl Acad Sci USA 103:18190–18195PubMedCrossRefGoogle Scholar
  30. Matsuda Y, Nishida-Umehara Ch, Tarui H, Kuroiwa A, Yamada K, Isobe T, Ando J, Fujiwara A, Hirao Y, Nishimura O, Ishijima J, Hayashi A, Saito T, Murakami T, Murakami Y, Kuratani S, Agata K (2005) Highly conserved linkage homology between birds and turtles: bird and turtle chromosomes are precise counterparts of each other. Chromosome Res 13:601–615PubMedCrossRefGoogle Scholar
  31. Mühlmann-Díaz MC, Ulsh BA, Whicker FW, Hinton TG, Congdon JD, Robinson JF, Bedford JS (2001) Conservation of chromosome 1 in turtles over 66 million years. Cytogenet Cell Genet 92:139–143PubMedCrossRefGoogle Scholar
  32. Nakatani Y, Takeda H, Kohara Y, Morishita S (2007) Reconstruction of the vertebrate ancestral genome reveals dynamic genome reorganization in early vertebrates. Genome Res 17:1254–1265PubMedCrossRefGoogle Scholar
  33. Nanda I, Haaf T, Schartl M, Schmid M, Burt DW (2002) Comparative mapping of Z-orthologous genes in vertebrates: implications for the evolution of avian sex chromosomes. Cytogenet Genome Res 99:178–184PubMedCrossRefGoogle Scholar
  34. Nishida-Umehara C, Tsuda Y, Ishijima J, Ando J, Fujiwara A, Matsuda Y, Griffin DK (2007) The molecular basis of chromosome orthologies and sex chromosomal differentiation in palaeognathous birds. Chromosome Res 15:721–734PubMedCrossRefGoogle Scholar
  35. O’Meally D, Patel HR, Stiglec R, Sarre SD, Geordes A, Graves JAM, Ezaz T (2010) Non-homologous sex chromosomes of birds and snakes share repetitive sequences. Chromosome Res 18:787–800PubMedCrossRefGoogle Scholar
  36. Olmo E (1986) A reptilia. In: John B (ed) Animal cytogenetics 4, Chordata 3. Gebrueder Borntraeger, Berlin, pp 1–100Google Scholar
  37. Olmo E (2008) Trends in the evolution of reptilian chromosomes. Integr Comp Biol 48:486–493PubMedCrossRefGoogle Scholar
  38. Organ C, Janes DE (2008) Evolution of sex chromosomes in Sauropsida. Integr Comp Biol 48:512–519PubMedCrossRefGoogle Scholar
  39. Pokorná M, Kratochvíl L (2009) Phylogeny of sex-dermining mechanisms in squamate reptiles: are sex chromosomes an evolutionary trap? Zool J Linn Soc 156:168–183CrossRefGoogle Scholar
  40. Pokorná M, Rábová M, Ráb P, Ferguson-Smith MA, Rens W, Kratochvíl L (2010) Differentiation of sex chromosomes and karyotypic evolution in the eye-lid geckos (Squamata: Gekkota: Eublepharidae), a group with different modes of sex determination. Chromosome Res 18:809–820PubMedCrossRefGoogle Scholar
  41. Ráb P, Rábová M, Pereira CS, Collares-Pereira MJ, Pelikánová Š (2008) Chromosome studies of European cyprinid fishes: interspecific homology of leuciscine cytotaxonomic marker—the largest subtelocentric chromosome pair as revelaed by cross-species painting. Chromosome Res 16:863–873PubMedCrossRefGoogle Scholar
  42. Rens W, O’Brien PCM, Yang F, Graves JAM, Ferguson-Smith MA (1999) Karyotype relationships between four distantly related marsupials revealed by reciprocal chromosome painting. Chromosome Res 7:461–474PubMedCrossRefGoogle Scholar
  43. Rens W, O’Brien PCM, Yang F, Solanky N, Perelman P, Graphodatsky AS, Ferguson MW, Svartman M, De Leo AA, Graves JAM, Ferguson-Smith MA (2001) Karyotype relationships between distantly related marsupials from South America and Australia. Chromosome Res 9:301–308PubMedCrossRefGoogle Scholar
  44. Rens W, O’Brien PCM, Fairclough H, Harman L, Graves JAM, Ferguson-Smith MA (2003) Reversal and convergence in marsupial chromosome evolution. Cytogenet Genome Res 102:282–290PubMedCrossRefGoogle Scholar
  45. Rens W, Fu B, O’Brien PCM, Ferguson-Smith MA (2006) Cross-species chromosome painting. Nat Protoc 1:783–790PubMedCrossRefGoogle Scholar
  46. Rens W, O’Brien PCM, Grützner F, Clarke O, Graphodatskaya D, Tsend-Ayush E, Trifonov VA, Skelton H, Wallis MC, Johnston S, Veyrunes F, Graves JAM, Ferguson-Smith MA (2007) The multiple sex chromosomes of platypus and echidna are not completely identical and several share homology with the avian Z. Genome Biol 8:R243PubMedCrossRefGoogle Scholar
  47. Seipp R, Henkel FW (2000) Rhacodactylus: biology, natural history and husbandry. Edition Chimaira, GermanyGoogle Scholar
  48. Shedlock AM, Edwards SV (2009) Amniota. In: Hedges SB, Kumar S (eds) The timetree of life. Oxford University Press, New York, pp 375–379Google Scholar
  49. Shetty S, Griffin DK, Graves JAM (1999) Comparative painting reveals strong chromosome homology over 80 million years of bird evolution. Chromosome Res 7:289–295PubMedCrossRefGoogle Scholar
  50. Shibaike Y, Takahashi Y, Arikura I, Iiizumi R, Kitakawa S, Sakai M, Imaoka C, Shiro H, Tanaka H, Akakubo N, Nakano M, Watanabe M, Ohne K, Kubota S, Kohno S, Ota H (2009) Chromosome evolution in the lizard genus Gekko (Gekkonidae, Squamata, Reptilia) in the east Asian islands. Cytogenet Genome Res 127:182–190PubMedCrossRefGoogle Scholar
  51. Smith JJ, Voss SR (2007) Bird and mammal sex chromosome orthologs map to the same autosomal region in salamander (Ambystoma). Genetics 177:607–613PubMedCrossRefGoogle Scholar
  52. Solleder E, Schmid M (1984) XX/XY-sex chromosomes in Gekko gecko (Sauria, Reptilia). Amphibia-Reptilia 5:339–345Google Scholar
  53. Srikulnath K, Nishida C, Matsubara K, Uno Y, Thongpan A, Suputtitada S, Apisitwanich S, Matsuda Y (2009) Karyotypic evolution in squamate reptiles: comparative gene mapping revealed highly conserved linkage homology between the butterfly lizard (Leiolepis reevesii rubritaeniata, Agamidae, Lacertilia) and the Japanese four-striped rat snake (Elaphe quadrivirgata, Colubridae, Serpentes). Chromosome Res 17:975–986PubMedCrossRefGoogle Scholar
  54. Telenius H, Pelmear AH, Tunnacliffe A, Carter NP, Behmel A, Ferguson-Smith MA, Nordenskjöld M, Pfragner R, Ponder BA (1992) Cytogenetic analysis by chromosome painting using DOP-PCR amplified flow-sorted chromosomes. Genes Chromosom Cancer 4:257–263PubMedCrossRefGoogle Scholar
  55. Veyrunes F, Waters Paul D, Miethke P, Rens W, McMillan D, Alsop AE, Frank G, Deakin JE, Whittington CM, Schatzkamer K, Kremitzki CL, Graves T, Ferguson-Smith MA, Warren W, Graves JAM (2008) Bird-like sex chromosomes of platypus imply recent origin of mammal sex chromosomes. Genome Res 18:965–973PubMedCrossRefGoogle Scholar
  56. Vidal N, Hedges SB (2005) The phylogeny of squamate reptiles (lizards, snakes, and amphisbaenians) inferred from nine nuclear protein-coding genes. C R Biol 328:1000–1008PubMedGoogle Scholar
  57. Yang F, Carter NP, Shi L, Ferguson-Smith MA (1995) A comparative study of karyotypes of muntjacs by chromosome painting. Chromosoma 103:642–652PubMedCrossRefGoogle Scholar
  58. Yang F, Alkalaeva EZ, Perelman PL, Pardini AT, Harrison WR, O’Brien PCM, Fu B, Graphodatsky AS, Ferguson-Smith MA, Robinson TJ (2003) Reciprocal chromosome painting among human, aardvark, and elephant (superorder Afrotheria) reveals the likely eutherian ancestral karyotype. Proc Natl Acad Sci USA 100:1062–1066PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Martina Pokorná
    • 1
    • 2
    • 5
  • Massimo Giovannotti
    • 3
    • 5
  • Lukáš Kratochvíl
    • 1
    Email author
  • Fumio Kasai
    • 5
  • Vladimir A. Trifonov
    • 4
    • 5
  • Patricia C. M. O’Brien
    • 5
  • Vincenzo Caputo
    • 3
  • Ettore Olmo
    • 3
  • Malcolm A. Ferguson-Smith
    • 5
  • Willem Rens
    • 5
    Email author
  1. 1.Faculty of ScienceCharles University in PraguePrague 2Czech Republic
  2. 2.Department of Vertebrate Evolutionary Biology and Genetics, Institute of Animal Physiology and GeneticsAcademy of Sciences of the Czech RepublicLiběchovCzech Republic
  3. 3.Dipartimento di Biochimica, Biologia e GeneticaUniversità Politecnica delle MarcheAnconaItaly
  4. 4.Institute of Chemical Biology and Fundamental Medicine SB RASNovosibirskRussia
  5. 5.Cambridge Resource Centre for Comparative Genomics, Department of Veterinary MedicineUniversity of CambridgeCambridgeUK

Personalised recommendations