Abstract
The genus Daphnia has an intriguing reproductive mode of cyclical parthenogenesis. This reproductive mode has been studied for centuries, but cytogenetic information is lacking due to technical limitations of classical methods. We have developed methods for the preparation and examination of meiotic chromosomes of Daphnia pulex from oocytes and spermatocytes. Oocyte chromosome preparations are obtained by isolating individual oocytes after the release of yolk granules from the ovary using pressure and capillary action. Spermatocyte chromosomes are prepared using a conventional squash method. Cryosectioning is an easy and fast way to prepare sections. We also illustrate the application of immunofluorescence staining against α tubulin, as well as fluorescence in situ hybridization (FISH) using the intergenic spacer of ribosomal DNA or single-copy cosmid clones.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
The URL is http://genome.jgi-psf.org/Dappu1/Dappu1.home.html
Zaffagnini, F. (1987) Reproduction in Daphnia, in Daphnia memorie (Peters, R.H. and de Bernardi, R., eds.) 45, pp. 245–284.
Hoskins, R.A., Carlson, J.W., Kennedy, C., Acevedo, D., Evans-Holm, M., Frise, E., Wan, K.H., Park, S., Mendez-Laqo, M., Rossi, F., Villasante, A., Dimitri, P., Karpen, G.H., and Celniker, S.E. (2007) Sequence finishing and mapping of Drosophila melanogaster heterochromatin. Science. 316, 1625–1628.
Fritsch, C., Ploeger, G., and Arndt-Jovin, D.J. (2006) Drosophila under the lens: imaging from chromosomes to whole embryos. Chromosome Res. 14, 451–464.
MacQueen, A.J., Phllips, C.M., Bhalla, N., Weiser, P., Villeneuve, A.M., and Dernburg, A.F. (2005) Chromosome sites play dual roles to establish homologous synapsis during meiosis in C. elegans. Cell. 123, 1037–1050.
Alpi, A., Pasierbek, P., Gartner, A., and Loidl, J. (2003) Genetic and cytological characterization of recombination protein RAD-51 in Caenorhabditis elegans. Chromosoma. 112, 6–16.
Ojima, Y. (1958) A cytological study on the development and maturation of the parthenogenetic and sexual eggs of Daphnia pulex (Crustacea-Cladocera). Kwansei Gakuin Unic. Ann. Studies. 6, 123–176.
Zaffagnini, F. and Sabelli, B. (1972) Karyologic observations on the maturation of the summer and winter eggs of Daphnia pulex and Daphnia middendorffiana. Chromosoma. 36, 193–203.
Acknowledgments
We would like to thank Armin Moczek and Teiya Kijimoto for their help with cryosectioning as well as helpful suggestions. This work was supported by National Science Foundation grant FIBR-0328516.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tsuchiya, D., Eads, B.D., Zolan, M.E. (2009). Methods for Meiotic Chromosome Preparation, Immunofluorescence, and Fluorescence in situ Hybridization in Daphnia pulex . In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 558. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-103-5_14
Download citation
DOI: https://doi.org/10.1007/978-1-60761-103-5_14
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-102-8
Online ISBN: 978-1-60761-103-5
eBook Packages: Springer Protocols