Advertisement

Chromosoma

, Volume 118, Issue 5, pp 575–589 | Cite as

Cohesin axis maturation and presence of RAD51 during first meiotic prophase in a true bug

  • Alberto Viera
  • Juan Luis Santos
  • María Teresa Parra
  • Adela Calvente
  • Rocío Gómez
  • Roberto de la Fuente
  • José Ángel Suja
  • Jesús Page
  • Julio S. RufasEmail author
Research Article

Abstract

We have analyzed in a true bug, Graphosoma italicum (Pentatomidae, Hemiptera), the temporal and functional relationships between recombination events, synapsis progression, and SMC1α and SMC3 cohesin axis maturation throughout the male first meiotic prophase. The localization of the histone variant histone H3 trimethylated at lysine 9 at chromosome ends has allowed us to determine the association of these heterochromatic domains through prophase I stages. Results highlighted that cohesins provide to be good markers for synapsis progression since the formation, morphology, and development of the SMC1α and SMC3 cohesin axes resemble the synaptonemal complex dynamics and, also, that in this species the initiation of recombination precedes synapsis. In addition, we have carried out an accurate cytological characterization of the diffuse stage, which takes place after pachytene, and also analyzed the presence of the cohesin subunits, SMC1α and SMC3, and the recombinase RAD51 at this stage. The mechanisms underlying the absence of SMC1α and SMC3 axes from the diffuse stage onwards are discussed.

Keywords

Seminiferous Tubule Synaptonemal Complex Diffuse Stage Sister Chromatid Cohesion Cohesin Complex 
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.

Notes

Acknowledgements

We would like to express our gratitude to Consejería de Medio Ambiente y Ordenación del Territorio (Comunidad de Madrid; España) for emitting the authorization for animal recollections in natural populations, to Dr. José Luis Barbero for generously supplying us the anti-SMC3 anti-SMC1α antibodies, to Dr. Kim McKim for his generous gift of the anti-H2Av antibody, and to Dr. García de la Verga for the critical reading of the manuscript. This work was supported by grants BFU2006-06655, BFU2005-02431, BFU200800459, and BFU2008-00300-BCM from the Ministerio de Educación y Ciencia Spain.

Supplementary material

Video 1

SMC3 distribution in squashed spermatocytes. 3-D reconstruction of a field of spermatocytes immunolabeled for SMC3 (green) and DAPI-stained (pseudocolored in red). Leptotene (Le), zygotene (Zy), early-, mid-, and late-pachytene (Pearly, Pmid, and Plate), diffuse, and metaphase I (MI) spermatocytes are observed and one spermatid (Sp). Note the presence of a brighter structure in pachytene and diffuse spermatocytes, corresponding to the chromatin of the sex chromosomes (XY). This video corresponds to the reconstructions of the spermatocytes shown in Fig. 1. See text for further details (MOV 710 kb)

412_2009_218_MOESM2_ESM.mov (3.1 mb)
Video 2 RAD51 distribution in a pachytene spermatocyte. 3-D reconstruction of a squashed pachytene spermatocyte immunolabeled for RAD51 (red) and DAPI-stained (blue). Despite RAD51 being located on the autosomal chromatin, it is noteworthy that sex chromosomes (XY) do not show RAD51 signals (MOV 3250 kb)

References

  1. Calvente A, Viera A, Page J, Parra MT, Gomez R, Suja JA, Rufas JS, Santos JL (2005) DNA double-strand breaks and homology search: inferences from a species with incomplete pairing and synapsis. J Cell Sci 118:2957–2963PubMedCrossRefGoogle Scholar
  2. Cardoso H, Stoll M, Dutra A, Oliver G, Di Tomaso MV (1986) Characterization of the diffuse stage in the male meiotic prophase and karyotype of Scapteriscus borellii (Orthoptera: Grylloidea). Genetica 71:23–29CrossRefGoogle Scholar
  3. Darlington CD, LaCour LF (1969) The handling of chromosomes. Allen and Unwin, LondonGoogle Scholar
  4. de la Fuente R, Parra MT, Viera A, Calvente A, Gomez R, Suja JA, Rufas JS, Page J (2007) Meiotic pairing and segregation of achiasmate sex chromosomes in eutherian mammals: the role of SYCP3 protein. PLoS Genet 3:e198PubMedCrossRefGoogle Scholar
  5. Dernburg AF, McDonald K, Moulder G, Barstead R, Dresser M, Villeneuve AM (1998) Meiotic recombination in C. elegans initiates by a conserved mechanism and is dispensable for homologous chromosome synapsis. Cell 94:387–398PubMedCrossRefGoogle Scholar
  6. Eijpe M, Heyting C, Gross B, Jessberger R (2000) Association of mammalian SMC1 and SMC3 proteins with meiotic chromosomes and synaptonemal complexes. J Cell Sci 113(Pt 4):673–682PubMedGoogle Scholar
  7. Esponda P, Stockert JC (1978) Localization of the synaptonemal complex under the light microscope. Chromosoma 68:83–90PubMedCrossRefGoogle Scholar
  8. Garcia M, Dietrich AJ, Freixa L, Vink AC, Ponsa M, Egozcue J (1987) Development of the first meiotic prophase stages in human fetal oocytes observed by light and electron microscopy. Hum Genet 77:223–232PubMedCrossRefGoogle Scholar
  9. Garcia-Cao M, O'Sullivan R, Peters AH, Jenuwein T, Blasco MA (2004) Epigenetic regulation of telomere length in mammalian cells by the Suv39h1 and Suv39h2 histone methyltransferases. Nat Genet 36:94–99PubMedCrossRefGoogle Scholar
  10. Grelon M, Vezon D, Gendrot G, Pelletier G (2001) AtSPO11-1 is necessary for efficient meiotic recombination in plants. Embo J 20:589–600PubMedCrossRefGoogle Scholar
  11. Henderson KA, Keeney S (2005) Synaptonemal complex formation: where does it start? Bioessays 27:995–998PubMedCrossRefGoogle Scholar
  12. Hodges CA, Revenkova E, Jessberger R, Hassold TJ, Hunt PA (2005) SMC1beta-deficient female mice provide evidence that cohesins are a missing link in age-related nondisjunction. Nat Genet 37:1351–1355PubMedCrossRefGoogle Scholar
  13. Jenuwein T, Allis CD (2001) Translating the histone code. Science 293:1074–1080PubMedCrossRefGoogle Scholar
  14. Kitajima TS, Yokobayashi S, Yamamoto M, Watanabe Y (2003) Distinct cohesin complexes organize meiotic chromosome domains. Science 300:1152–1155PubMedCrossRefGoogle Scholar
  15. Klásterská I (1976) A new look on the role of the diffuse stage in problems of plant and animal species. Hereditas 82:193–204CrossRefGoogle Scholar
  16. Klásterská I (1977) The concept of the prophase on meiosis. Hereditas 86:205–210CrossRefGoogle Scholar
  17. Kleckner N (1996) Meiosis: how could it work? Proc Natl Acad Sci U S A 93:8167–8174PubMedCrossRefGoogle Scholar
  18. Mahadevaiah SK, Turner JM, Baudat F, Rogakou EP, de Boer P, Blanco-Rodriguez J, Jasin M, Keeney S, Bonner WM, Burgoyne PS (2001) Recombinational DNA double-strand breaks in mice precede synapsis. Nat Genet 27:271–276PubMedCrossRefGoogle Scholar
  19. McKim KS, Hayashi-Hagihara A (1998) mei-W68 in Drosophila melanogaster encodes a Spo11 homolog: evidence that the mechanism for initiating meiotic recombination is conserved. Genes Dev 12:2932–2942PubMedCrossRefGoogle Scholar
  20. Nicklas RB, Kubai DF, Hays TS (1982) Spindle microtubules and their mechanical associations after micromanipulation in anaphase. J Cell Biol 95:91–104PubMedCrossRefGoogle Scholar
  21. Page SL, Hawley RS (2004) The genetics and molecular biology of the synaptonemal complex. Annu Rev Cell Dev Biol 20:525–558PubMedCrossRefGoogle Scholar
  22. Page J, Suja JA, Santos JL, Rufas JS (1998) Squash procedure for protein immunolocalization in meiotic cells. Chromosome Res 6:639–642PubMedCrossRefGoogle Scholar
  23. Page J, Berrios S, Rufas JS, Parra MT, Suja JA, Heyting C, Fernandez-Donoso R (2003) The pairing of X and Y chromosomes during meiotic prophase in the marsupial species Thylamys elegans is maintained by a dense plate developed from their axial elements. J Cell Sci 116:551–560PubMedCrossRefGoogle Scholar
  24. Page J, de la Fuente R, Gomez R, Calvente A, Viera A, Parra MT, Santos JL, Berrios S, Fernandez-Donoso R, Suja JA et al (2006) Sex chromosomes, synapsis, and cohesins: a complex affair. Chromosoma 115:250–259PubMedCrossRefGoogle Scholar
  25. Parra MT, Page J, Yen TJ, He D, Valdeolmillos A, Rufas JS, Suja JA (2002) Expression and behaviour of CENP-E at kinetochores during mouse spermatogenesis. Chromosoma 111:53–61PubMedCrossRefGoogle Scholar
  26. Parra MT, Viera A, Gomez R, Page J, Benavente R, Santos JL, Rufas JS, Suja JA (2004) Involvement of the cohesin Rad21 and SCP3 in monopolar attachment of sister kinetochores during mouse meiosis I. J Cell Sci 117:1221–1234PubMedCrossRefGoogle Scholar
  27. Pelttari J, Hoja MR, Yuan L, Liu JG, Brundell E, Moens P, Santucci-Darmanin S, Jessberger R, Barbero JL, Heyting C et al (2001) A meiotic chromosomal core consisting of cohesin complex proteins recruits DNA recombination proteins and promotes synapsis in the absence of an axial element in mammalian meiotic cells. Mol Cell Biol 21:5667–5677PubMedCrossRefGoogle Scholar
  28. Peters AH, Plug AW, van Vugt MJ, de Boer P (1997) A drying-down technique for the spreading of mammalian meiocytes from the male and female germ line. Chromosome Res 5:66–68PubMedCrossRefGoogle Scholar
  29. Peters AH, O'Carroll D, Scherthan H, Mechtler K, Sauer S, Schofer C, Weipoltshammer K, Pagani M, Lachner M, Kohlmaier A et al (2001) Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell 107:323–337PubMedCrossRefGoogle Scholar
  30. Pigozzi MI, Solari AJ (2003) Differential immunolocalization of a putative Rec8p in meiotic autosomes and sex chromosomes of triatomine bugs. Chromosoma 112:38–47PubMedCrossRefGoogle Scholar
  31. Prieto I, Tease C, Pezzi N, Buesa JM, Ortega S, Kremer L, Martinez A, Martinez AC, Hulten MA, Barbero JL (2004) Cohesin component dynamics during meiotic prophase I in mammalian oocytes. Chromosome Res 12:197–213PubMedCrossRefGoogle Scholar
  32. Revenkova E, Jessberger R (2005) Keeping sister chromatids together: cohesins in meiosis. Reproduction 130:783–790PubMedCrossRefGoogle Scholar
  33. Revenkova E, Eijpe M, Heyting C, Gross B, Jessberger R (2001) Novel meiosis-specific isoform of mammalian SMC1. Mol Cell Biol 21:6984–6998PubMedCrossRefGoogle Scholar
  34. Roeder GS (1997) Meiotic chromosomes: it takes two to tango. Genes Dev 11:2600–2621PubMedCrossRefGoogle Scholar
  35. Rufas JS, Gimenez-Martin G (1986) Ultrastructure of the kinetochore in Graphosoma italicum (Hemiptera, Heteroptera). Protoplasma 132:142–148CrossRefGoogle Scholar
  36. Stockert JC (1977) Osmium tetroxide/p-phenylenediamine staining of nucleoli and Balbiani rings in Chironomus salivary glands. Histochemistry 53:43–56PubMedCrossRefGoogle Scholar
  37. Storlazzi A, Tesse S, Ruprich-Robert G, Gargano S, Poggeler S, Kleckner N, Zickler D (2008) Coupling meiotic chromosome axis integrity to recombination. Genes Dev 22:796–809PubMedCrossRefGoogle Scholar
  38. Strahl BD, Allis CD (2000) The language of covalent histone modifications. Nature 403:41–45PubMedCrossRefGoogle Scholar
  39. Suja JA, del Cerro AL, Page J, Rufas JS, Santos JL (2000) Meiotic sister chromatid cohesion in holocentric sex chromosomes of three heteropteran species is maintained in absence of axial elements. Chromosoma 109:35–43PubMedCrossRefGoogle Scholar
  40. Tesse S, Storlazzi A, Kleckner N, Gargano S, Zickler D (2003) Localization and roles of Ski8p protein in Sordaria meiosis and delineation of three mechanistically distinct steps of meiotic homolog juxtaposition. Proc Natl Acad Sci U S A 100:12865–12870PubMedCrossRefGoogle Scholar
  41. Valdeolmillos AM, Viera A, Page J, Prieto I, Santos JL, Parra MT, Heck MM, Martinez AC, Barbero JL, Suja JA et al (2007) Sequential loading of cohesin subunits during the first meiotic prophase of grasshoppers. PLoS Genet 3:e28PubMedCrossRefGoogle Scholar
  42. Viera A, Parra MT, Page J, Santos JL, Rufas JS, Suja JA (2003) Dynamic relocation of telomere complexes in mouse meiotic chromosomes. Chromosome Res 11:797–807PubMedCrossRefGoogle Scholar
  43. Viera A, Calvente A, Page J, Parra MT, Gomez R, Suja JA, Rufas JS, Santos JL (2004a) X and B chromosomes display similar meiotic characteristics in male grasshoppers. Cytogenet Genome Res 106:302–308PubMedCrossRefGoogle Scholar
  44. Viera A, Santos JL, Page J, Parra MT, Calvente A, Cifuentes M, Gomez R, Lira R, Suja JA, Rufas JS (2004b) DNA double-strand breaks, recombination and synapsis: the timing of meiosis differs in grasshoppers and flies. EMBO Rep 5:385–391PubMedCrossRefGoogle Scholar
  45. Viera A, Page J, Rufas JS (2009a) Inverted meiosis: the true bugs as a model to study. Genome Dyn 5:137–156PubMedCrossRefGoogle Scholar
  46. Viera A, Santos JL, Rufas JS (2009b) Relationship between incomplete synapsis and chiasma localization. Chromosoma 118:377–389PubMedCrossRefGoogle Scholar
  47. Wilson EB (1928) The cell in development and heredity. Macmillan, New YorkGoogle Scholar
  48. Zickler D, Kleckner N (1999) Meiotic chromosomes: integrating structure and function. Annu Rev Genet 33:603–754PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Alberto Viera
    • 1
  • Juan Luis Santos
    • 2
  • María Teresa Parra
    • 1
  • Adela Calvente
    • 1
  • Rocío Gómez
    • 1
  • Roberto de la Fuente
    • 1
  • José Ángel Suja
    • 1
  • Jesús Page
    • 1
  • Julio S. Rufas
    • 1
    • 3
    Email author
  1. 1.Departamento de Biología, Edificio de Biológicas, Facultad de CienciasUniversidad Autónoma de MadridMadridSpain
  2. 2.Departamento de Genética, Facultad de BiologíaUniversidad Complutense de MadridMadridSpain
  3. 3.Unidad de Biología Celular, Departamento de Biología, Facultad de Ciencias, Edificio de Ciencias BiológicasUniversidad Autónoma de MadridMadridSpain

Personalised recommendations