Chromosome Structure and Homologous Chromosome Association During Meiotic Prophase in Caenorhabditiselegans

  • Kentaro NabeshimaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 745)


Successful meiotic recombination is driven by a series of programmed chromosome dynamics that include changes in the protein composition of meiotic chromosomes and the juxtaposition of homologous chromosomes. The simultaneous visualization of both chromosome-bound proteins and the status of homologous association is an important experimental approach to analyze the mechanisms supporting proper meiotic chromosome association. One of a number of model organisms used for meiosis research, the nematode Caenorhabditis elegans offers an excellent environment to study meiotic chromosome dynamics. Here I will describe how to visualize both chromosome structure and specific chromosomal loci simultaneously, in a whole-mount C. elegans germ line. It combines immunofluorescent (IF) staining for a meiotic chromosome structural component with fluorescent in situ hybridization (FISH).

Key words

C. elegans chromosome axis FISH germ line homologous pairing immunofluorescence meiosis synaptonemal complex whole-mount gonad 



The author would like to thank Dr. Anne M. Villeneuve for extensive support and helpful suggestions during part of the development of this method as well as for providing anti-SYP-1 antibody and Dr. Raymond Chan for critical reading of the manuscript and insightful comments. This work was supported by March of Dimes, Basil O’Conner Starter Scholar Award (#5-FY07-666).


  1. 1.
    Moens, P.B., and Pearlman, R.E. (1988) Chromatin organization at meiosis. Bioessays 9, 151–153.PubMedCrossRefGoogle Scholar
  2. 2.
    von Wettstein, D., Rasmussen, S.W., and Holm, P.B. (1984) The synaptonemal complex in genetic segregation. Ann Rev Genet 18, 331–413.CrossRefGoogle Scholar
  3. 3.
    Zetka, M.C., Kawasaki, I., Strome, S., and Muller, F. (1999) Synapsis and chiasma formation in Caenorhabditis elegans require HIM-3, a meiotic chromosome core component that functions in chromosome segregation. Genes Dev 13, 2258–2270.PubMedCrossRefGoogle Scholar
  4. 4.
    Martinez-Perez, E., and Villeneuve, A.M. (2005) HTP-1-dependent constraints coordinate homolog pairing and synapsis and promote chiasma formation during C. elegans meiosis. Genes Dev 19, 2727–2743.PubMedCrossRefGoogle Scholar
  5. 5.
    Couteau, F., and Zetka, M. (2005) HTP-1 coordinates synaptonemal complex assembly with homolog alignment during meiosis in C. elegans. Genes Dev 19, 2744–2756.PubMedCrossRefGoogle Scholar
  6. 6.
    Goodyer, W., Kaitna, S., Couteau, F., Ward, J.D., Boulton, S.J., and Zetka, M. (2008) HTP-3 Links DSB formation with homolog pairing and crossing over during C. elegans meiosis. Dev Cell 14, 263–274.PubMedCrossRefGoogle Scholar
  7. 7.
    Pasierbek, P., Jantsch, M., Melcher, M., Schleiffer, A., Schweizer, D., and Loidl, J. (2001) A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction. Genes Dev 15, 1349–1360.PubMedCrossRefGoogle Scholar
  8. 8.
    MacQueen, A.J., Colaiacovo, M.P., McDonald, K., and Villeneuve, A.M. (2002) Synapsis-dependent and -independent mechanisms stabilize homolog pairing during meiotic prophase in C. elegans. Genes Dev 16, 2428–2442.PubMedCrossRefGoogle Scholar
  9. 9.
    Colaiacovo, M.P., MacQueen, A.J., Martinez-Perez, E., McDonald, K., Adamo, A., La Volpe, A., and Villeneuve, A.M. (2003) Synaptonemal complex assembly in C. elegans is dispensable for loading strand-exchange proteins but critical for proper completion of recombination. Dev Cell 5, 463–474.PubMedCrossRefGoogle Scholar
  10. 10.
    Smolikov, S., Eizinger, A., Schild-Prufert, K., Hurlburt, A., McDonald, K., Engebrecht, J., Villeneuve, A.M., and Colaiacovo, M.P. (2007) SYP-3 restricts synaptonemal complex assembly to bridge paired chromosome axes during meiosis in Caenorhabditis elegans. Genetics 176, 2015–2025.PubMedCrossRefGoogle Scholar
  11. 11.
    Smolikov, S., Schild-Prufert, K., and Colaiacovo, M.P. (2009) A yeast two-hybrid screen for SYP-3 interactors identifies SYP-4, a component required for synaptonemal complex assembly and chiasma formation in Caenorhabditis elegans meiosis. PLoS Genet 5, e1000669.PubMedCrossRefGoogle Scholar
  12. 12.
    Couteau, F., Nabeshima, K., Villeneuve, A., and Zetka, M. (2004) A component of C. elegans meiotic chromosome axes at the interface of homolog alignment, synapsis, nuclear reorganization, and recombination. Curr Biol 14, 585–592.PubMedCrossRefGoogle Scholar
  13. 13.
    Nabeshima, K., Villeneuve, A.M., and Hillers, K.J. (2004) Chromosome-wide regulation of meiotic crossover formation in Caenorhabditis elegans requires properly assembled chromosome axes. Genetics 168, 1275–1292.PubMedCrossRefGoogle Scholar
  14. 14.
    Coulson, A., Waterston, R., Kiff, J., Sulston, J., and Kohara, Y. (1988) Genome linking with yeast artificial chromosomes. Nature 335, 184–186.PubMedCrossRefGoogle Scholar
  15. 15.
    Edgar, L.G. (1995) Blastomere culture and analysis. In Methods in cell biology: Caenorhabditis elegans, modern biological analysis of an organism, H.F. Epstein and D.C. Shakes, eds. (New York, NY: Academic Press), p. 317.Google Scholar
  16. 16.
    Roohi, J., Cammer, M., Montagna, C., and Hatchwell, E. (2008) An improved method for generating BAC DNA suitable for FISH. Cytogenetic Genome Res 121, 7–9.CrossRefGoogle Scholar
  17. 17.
    Dernburg, A.F. (1999) Fluorescence in situ hybridization in whole-mount tissues. In Chromosome structural analysis, a practical approach, W.A. Bickmore, ed. (Oxford: Oxford University Press), p. 142.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Cell and Developmental BiologyUniversity of Michigan, Medical SchoolAnn ArborUSA

Personalised recommendations