Conservation Genetics

, Volume 12, Issue 5, pp 1159–1171 | Cite as

Patterns of variability at the major histocompatibility class I and class II loci in populations of the endangered cyprinid Ladigesocypris ghigii

  • Katerina A. MoutouEmail author
  • Zissis Mamuris
  • Tania Firme
  • Maria Kontou
  • Theologia Sarafidou
  • Maria Th. Stoumboudi
Research Article


The patterns of MHC diversity were studied at UAA and DAB1 loci and the two domains involved in the recognition of antigenic peptides (α2 and β1, respectively) in eight Ladigesocypris ghigii populations inhabiting streams and a concrete reservoir, in order to understand the significance of these genes in bottlenecked populations of an endemic species and develop conservation rationale. In agreement with previous study employing RAPD and mtDNA markers (Mamuris et al., Freshw Biol 50:1441–1453, 2005), both loci exhibited a very low level of polymorphism with only two and four alleles detected for UAA and DAB1, respectively. The functional MHC diversity was even lower since UAA alleles were distinguished by a single synonymous substitution. The type of habitat did not affect the level of polymorphism. Our data suggest that DAB1 polymorphism might be the outcome of the positive selection, imposed by the temporal and spatial variation of pathogen load, and the genetic drift as a result of successive habitat shrinkage and deterioration by water abstraction year after year. The populations studied were significantly less diverged at MHC loci than expected based on nuclear and mtDNA markers, suggesting that common parasites might act as causative factors to homogenize selection. Sufficient epidemiological data are required for the interpretation of the results and decision-making on suitable conservation actions.


Ladigesocypris ghigii MHC diversity UAA DAB1 Conservation 



The present study was funded by the Program of Postgraduate Studies “Biotechnology—Quality Assessment in Nutrition and the Environment”. Sample collection was funded by LIFE-Nature project B4-3200/98/445. The authors are indebted to Mr Costas Stamatis for invaluable technical assistance and to the two anonymous reviewers for their constructive comments.


  1. Aguilar A, Roemer G, Debenham S, Binns M, Garcelon D, Wayne RK (2004) High MHC diversity maintained by balancing selection in an otherwise genetically monomorphic mammal. Proc Natl Acad Sci 101:3490–3494PubMedCrossRefGoogle Scholar
  2. Apanius V, Penn D, Slev PR, Ruff LR, Potts WK (1997) The nature of selection on the major histocompatibility complex. Crit Rev Immunol 17:179–224PubMedGoogle Scholar
  3. Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modeling. Bioinformatics 22:195–201PubMedCrossRefGoogle Scholar
  4. Avise JC (2000) Phylogeography. Harvard University Press, CambridgeGoogle Scholar
  5. Babik W, Durka W, Radwan J (2005) Sequence diversity of the MHC DRB gene in the Eurasian beaver (Castor fiber). Mol Ecol 14:4249–4257PubMedCrossRefGoogle Scholar
  6. Bernatchez L, Landry C (2003) MHC studies in non-model vertebrates: what have we learned about natural selection in 15 years? J Evol Biol 16:363–377PubMedCrossRefGoogle Scholar
  7. Bingulac-Popovic J, Figueroa F, Sato A, Talbot WS, Johnson SL, Gates M, Postlethwait JH, Klein J (1997) Mapping of Mhc class I and class II regions to different linkage groups in the zebrafish, Danio rerio. Immunogenetics 46:129–134PubMedCrossRefGoogle Scholar
  8. Bolmer JL, Vargas FH, Parker PG (2007) Low MHC variation in the endangered Galápagos penguin (Spheniscus mendiculus). Immunogenetics 59:593–602CrossRefGoogle Scholar
  9. Brook BW, Tonkyn DW, O’Grady JJ, Frankham R (2002) Contribution of inbreeding to extinction risk in threatened species. Ecol Soc 6: article 16Google Scholar
  10. Brown JH, Jardetzky TS, Gorga JC, Stern LJ, Urban RG, Strominger JL, Wiley DC (1993) Three-dimensional structure of the human class II histocompatibility antigen HLA-DR1. Nature 364:33–39PubMedCrossRefGoogle Scholar
  11. Cárdenas C, Ortiz M, Balbín A, Villaveces JL, Patarroyo ME (2005) Allele effects in MHC-peptide interactions: a theoretical analysis of HLA-DRβ1*0101-HA and HLA-DRβ1*0401-HA complexes. Biochem Biophys Res Commun 330:1162–1167PubMedCrossRefGoogle Scholar
  12. Consuegra S, Megens HJ, Leon K, Stet RJM, Jordan WC (2005a) Patterns of variability at the major histocompatibility class II alpha locus in Atlantic salmon contrast with those at the class I locus. Immunogenetics 57:16–24PubMedCrossRefGoogle Scholar
  13. Consuegra S, Megens HJ, Schaschl H, Leon K, Stet RJM, Jordan WC (2005b) Rapid evolution of the MH class I locus results in different allelic compositions in recently diberged populations of Atlantic salmon. Mol Biol Evol 22:1095–1106PubMedCrossRefGoogle Scholar
  14. Corsini M (1995) Distribution of the populations of an endemic cyprinid (Ladigesocypris ghigii) in the island of Rhodes (Greece) and efforts for its conservation. Memoires de l’Institut Oceanographique, Leipzig, pp 63–67Google Scholar
  15. Dionne M, Miller KM, Dodson JJ, Caron F, Bernatchez L (2007) Clinal variation in MHC diversity with temperature: evidence for the role of host-pathogen interaction on local adaptation in Atlantic salmon. Evolution 61:2154–2164PubMedCrossRefGoogle Scholar
  16. Dixon B, Nagelkerke LAJ, Sibbing FA, Egberts E, Stet RJM (1996) Evolution of MHC class II β chain-encoding genes in the Lake Tana barbell species flock (Barbus intermedius complex). Immunogenetics 44:419–431PubMedGoogle Scholar
  17. Economidis PS (1995) Endangered freshwater fishes of Greece. Biol Conserv 72:201–211CrossRefGoogle Scholar
  18. Edwards SV, Hedrick PW (1998) Evolution and ecology of MHC molecules: from genomics to sexual selection. Trends Ecol Evol 13:305–311PubMedCrossRefGoogle Scholar
  19. Eizaguirre C, Lenz TL, Sommerfeld RD, Harrod C, Kalbe M, Milinski M (2010) Parasite diversity, patterns of MHC II variation and olfactory based mate choice in diverging tree-spined stickleback ecotypes. Evol Ecol 25:605–622CrossRefGoogle Scholar
  20. Ellegren H, Hartman G, Johansson M, Andersson L (1993) Major histocompatibility complex monomorphism and low levels of DNA fingerprinting variability in a reintroduced and rapidly expanding population of beavers. Proc Natl Acad Sci 90:8150–8153PubMedCrossRefGoogle Scholar
  21. Frankel OH, Soulé ME (1981) Conservation and evolution. Cambridge University Press, CambridgeGoogle Scholar
  22. Goudet J (2001) FSTAT. A program to estimate and test gene diversities and fixation indices (version 2.9.3). Updated from Goudet (1995)
  23. Graser R, Vincek V, Takami K, Klein J (1998) Analysis of zebrafish Mhc using BAC clones. Immunogenetics 47:318–325PubMedCrossRefGoogle Scholar
  24. Grimholt U, Larsen S, Nordmo R, Midtlyng P, Kjoeglum S, Storset A, Saebø S, Stet RJM (2003) MHC polymorphism and disease resistance in Atlantic salmon (Salmo salar); facing pathogens with single expressed major histocompatibility class I and class II loci. Immunogenetics 55:210–219PubMedCrossRefGoogle Scholar
  25. Hashimoto K, Nakanishi T, Kurosawa Y (1990) Isolation of carp genes encoding major histocompatibility complex antigens. Proc Natl Acad Sci 87:6863–6867PubMedCrossRefGoogle Scholar
  26. Hedrick PW (1994) Evolutionary genetics at the major histocompatibility complex. Am Nat 143:945–964CrossRefGoogle Scholar
  27. Hedrick PW (2002) Pathogen resistance and genetic variation at MHC loci. Evolution 56:1902–1908PubMedGoogle Scholar
  28. Hedrick PW, Parker KM, Gutiérrez-Espeleta GA, Rattink A, Lievers K (2000) Major histocompatibility complex variation in the Arabian Oryx. Evolution 5:2145–2151Google Scholar
  29. Hess CM, Edwards SV (2002) The evolution of the major histocompatibility complex in birds. Bioscience 52:423–431CrossRefGoogle Scholar
  30. Hodder KH, Bullock JM (1997) Translocations of native species in the UK: implications for biodiversity. J Appl Ecol 34:547–564CrossRefGoogle Scholar
  31. Hughes AL (1991) MHC polymorphism and the design of captive breeding programs. Conserv Biol 5:249–251CrossRefGoogle Scholar
  32. Jukes TH, Cantor CR (1969) Evolution of protein molecules. In: Munro HN (ed) Mammalian protein metabolism. Academic Press, New York, pp 21–132Google Scholar
  33. Kaufman J, Salomonsen J, Flajnik M (1994) Evolutionary conservation of MHC class I and class II molecules—different yet the same. Semin Immunol 6:411–424PubMedCrossRefGoogle Scholar
  34. Klein J (1986) Natural history of the major histocompatibility complex. Wiley, New YorkGoogle Scholar
  35. Klein J, Satta Y, O’hUigin C, Takahata N (1993) The molecular descent of the major histocompatibility complex. Annu Rev Immunol 11:269–295PubMedCrossRefGoogle Scholar
  36. Kottelat M (1997) European freshwater fishes—an heuristic checklist of the freshwater fishes of Europe (exclusive of former USSR), with an introduction for non-systematists and comments on nomenclature and conservation. Biol Bratisl Zool 52(Suppl 5):1–271Google Scholar
  37. Kraulis PJ (1991) Molscript—a program to produce both detailed and schematic plots of protein structures. J Appl Crystallogr 24:946–950CrossRefGoogle Scholar
  38. Kruiswijk CP, Hermsen T, Westphal AH, Savelkoul HFJ, Stet RJM (2002) A novel functional class I lineage in zebrafish (Danio rerio), carp (Cyprinus carpio) and large barbus (Barbus intermedius) showing an unusual conservation of the peptide binding domains. J Immunol 169:1936–1947PubMedGoogle Scholar
  39. Kruiswijk CP, Hermsen T, Fujiki K, Dixon B, Savelkoul HFJ, Stet RJM (2004) Analysis of genomic and expressed major histocompatibility class Ia and class II genes in a hexaploid Lake Tana African “large” barb individual (Barbus intermedius). Immunogenetics 55:770–781PubMedCrossRefGoogle Scholar
  40. Kruiswijk CP, Hermsen T, van Heerwaarden J, Dixon B, Savelkoul HFJ, Stet RJM (2005) Major histocompatibility genes in the Lake Tana African large barb species flock: evidence for complete partitioning of class II B, but not class I, genes among different species. Immunogenetics 56:894–908PubMedCrossRefGoogle Scholar
  41. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163PubMedCrossRefGoogle Scholar
  42. Lowe A, Harris S, Ashton P (2004) Ecological genetics: design, analysis, and application. Blackwell, OxfordGoogle Scholar
  43. Mamuris Z, Stoumboudi MTh, Stamatis C, Barbieri R, Moutou KA (2005) Genetic variation in population of the endangered fish Ladigesocypris ghigii and its implications for conservation. Freshw Biol 50:1441–1453CrossRefGoogle Scholar
  44. McFarland BJ, Beeson C (2002) Binding interactions between peptides and proteins of the class II major histocompatibility complex. Med Res Rev 22:168–203PubMedCrossRefGoogle Scholar
  45. Merritt EA, Bacon DJ (1997) Raster3D: photorealistic molecular graphics. Methods Enzymol 277:505–524PubMedCrossRefGoogle Scholar
  46. Miller PS, Hedrick PW (1991) MHC polymorphism and the design of captive breeding programs: simple solutions are not the answer. Conserv Biol 5:556–558CrossRefGoogle Scholar
  47. Miller HC, Lambert DM (2004) Genetic drift outweighs balancing selection in shaping post-bottleneck major histocompatibility complex variation in New Zealand robins (Petroicidae). Mol Ecol 13:3709–3721PubMedCrossRefGoogle Scholar
  48. Moravec F (2006) Systemic status of Rhabdochona leucaspii Kritscher, 1979 (Nematoda: Rhabdochonidae). Folia Parasitol 53:240PubMedGoogle Scholar
  49. Munguia-Vega A, Esquer-Garrigos Y, Rojas-Bracho L, Vazquez-Juarez R, Castro-Prieto A, Flores-Ramirez S (2007) Genetic drift vs natural selection in a long-term small isolated population: major histocompatibility complex class II variation in the Gulf of California endemic porpoise (Phocoena sinus). Mol Ecol 16:4051–4065PubMedCrossRefGoogle Scholar
  50. Nei M, Gojobori T (1986) Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol Biol Evol 3:418–426PubMedGoogle Scholar
  51. Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, LondonGoogle Scholar
  52. Nicholls A, Honig B (1991) A rapid finite-difference algorithm, utilizing successive over-relaxation to solve the Poisson-Boltzmann equation. J Comput Chem 12:435–445CrossRefGoogle Scholar
  53. Nielsen R (2005) Molecular signatures of natural selection. Annu Rev Genet 39:197–218PubMedCrossRefGoogle Scholar
  54. Ottová E, Šimková A, Martin J-F, de Bellocq JG, Gelnar M, Allienne J-F, Morand S (2005) Evolution and trans-species polymorphism of MHC class IIβ genes in cyprinid fish. Fish Shellfish Immunol 18:199–222PubMedCrossRefGoogle Scholar
  55. Peitsch MC (1995) Protein modeling by e-mail. Bio-Technology 13:658–660Google Scholar
  56. Ploegh H, Watts C (1998) Antigen recognition. Curr Opin Immunol 10:57–58PubMedCrossRefGoogle Scholar
  57. Potts WK, Wakeland EK (1993) Evolution of MHC genetic diversity: a tale of incest, pestilence and sexual preference. Trends Genet 9:408–412PubMedCrossRefGoogle Scholar
  58. Rammensee HG, Friede T, Stevanovic S (1995) MHC ligands and peptide motifs: first listing. Immunogenetics 41:178–228PubMedCrossRefGoogle Scholar
  59. Raymond M, Rousset F (1995) GENEPOP: population genetics software for exact tests and ecumenism. J Hered 86:248–249Google Scholar
  60. Richardson DS, Westerdahl H (2003) MHC diversity in two Acrocephalus species: the outbred great reed warbler and the inbred Seychelles warbler. Mol Ecol 12:3523–3529PubMedCrossRefGoogle Scholar
  61. Sato A, Figueroa F, Murray BW, Malaga-Trillo E, Zaleska-Rutczynska Z, Sultmann H, Toyosawa S, Wedekind C, Steck N, Klein J (2000) Nonlinkage of major histocompatibility complex class I and class II loci in bony fishes. Immunogenetics 51:108–116PubMedCrossRefGoogle Scholar
  62. Seddon JM, Baverstock PR (1999) Variation on islands: major histocompatibility complex (Mhc) polymorphism in populations of the Australian bush rat. Mol Ecol 8:2071–2079PubMedCrossRefGoogle Scholar
  63. Siebold C, Hansen BE, Wyer JR, Harlos K, Esnouf RE, Svejgaard A, Bell JI, Strominger JL, Jones EY, Fugger L (2004) Crystal structure of HLA-DQ0602 that protects against type 1 diabetes and confers strong susceptibility to narcolepsy. Proc Natl Acad Sci 101:1999–2004PubMedCrossRefGoogle Scholar
  64. Šimková A, Ottová E, Morand S (2006) MHC variability, life-traits and parasite diversity of European cyprinid fish. Evol Ecol 20:465–477Google Scholar
  65. Slade RW (1992) Limited MHC polymorphism in the southern elephant seal: implications for MHC evolution and marine mammal population biology. Proc R Soc B 249:163–171PubMedCrossRefGoogle Scholar
  66. Sommer S (2005) The importance of immune gene variability (MHC) in evolutionary ecology and conservation. Front Zool 2:16–34PubMedCrossRefGoogle Scholar
  67. Stern LJ, Brown JH, Jardetzky TS, Gorga JC, Urban RG, Strominger JL, Wiley DC (1994) Crystal-structure of the human class-II MHC protein HLA-DR1 complexed with an influenza-virus peptide. Nature 368:215–221PubMedCrossRefGoogle Scholar
  68. Stoumboudi MTh, Cowx IG (2003) Action plan for the endangered fish gizani (Ladigesocypris ghigii), endemic to Rhodes Island. Hellenic Centre for Marine Research, AthensGoogle Scholar
  69. Stoumboudi MTh, Barbieri R, Mamuris Z, Corsini-Foka MJ, Economou AN (2002a) Threatened fishes of the world: Ladigesocypris ghigii (Gianferrari, 1927) (Cyprinidae). Environ Biol Fish 65:340CrossRefGoogle Scholar
  70. Stoumboudi MTh, Barbieri R, Corsini-Foka MJ, Economou AN, Economidis PS (2002b) Aspects of the reproduction and early life history of Ladigesocypris ghigii a freshwater fish species endemic to Rhodes island (Greece): implementation to conservation. In: Collares-Perreira MJ, Cowx IG, Coelho MM (eds) Conservation of freshwater fishes: options for the future. Fishing News Books, Blackwell Science, Oxford, pp 178–185Google Scholar
  71. Takami K, Zaleska-Rutczynska Z, Figueroa F, Klein J (1997) Linkage of LMP, TAP, and RING3 with Mhc class I rather than class II genes in the zebrafish. J Immunol 159:6052–6060PubMedGoogle Scholar
  72. Takeuchi H, Figueroa F, O’hUigin C, Klein J (1995) Cloning and characterization of class I Mhc genes of the zebrafish, Brachydanio rerio. Immunogenetics 42:77–84PubMedCrossRefGoogle Scholar
  73. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4879–4882CrossRefGoogle Scholar
  74. Van Erp SHM, Dixon B, Figueroa F, Egberts E, Stet RJM (1996) Identification and characterization of the new major histocompatibility complex class I gene in carp (Cyprinus carpio L.). Immunogenetics 44:49–61PubMedCrossRefGoogle Scholar
  75. Vrijenhoek RC, Leberg PL (1991) Let’s not throw out the baby with the bathwater: a comment on management for MHC diversity in captive populations. Conserv Biol 5:252–254CrossRefGoogle Scholar
  76. Weber DS, Stewart BS, Schienman J, Lehman N (2004) Major histocompatibility complex variation at three class II loci in the northern elephant seal. Mol Ecol 13:711–718PubMedCrossRefGoogle Scholar
  77. Wegner KM, Reusch TBH, Kalbe M (2003) Multiple parasites are driving major histocompatibility complex polymorphism in the wild. J Evol Biol 16:224–232PubMedCrossRefGoogle Scholar
  78. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Katerina A. Moutou
    • 1
    Email author
  • Zissis Mamuris
    • 1
  • Tania Firme
    • 1
  • Maria Kontou
    • 1
  • Theologia Sarafidou
    • 1
  • Maria Th. Stoumboudi
    • 2
  1. 1.Department of Biochemistry and BiotechnologyUniversity of ThessalyLarissaGreece
  2. 2.Institute of Inland WatersHellenic Centre for Marine ResearchAnavyssosGreece

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