Chromosome Structure and Homologous Chromosome Association During Meiotic Prophase in Caenorhabditiselegans
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 wordsC. 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).
- 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.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
- 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
- 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