Skip to main content
SpringerLink
Log in

Cytological Analysis of Meiosis in Fixed Drosophila Ovaries

  • Protocol
  • First Online:
Book cover Meiosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 558))

Abstract

Methods are described to analyze two different parts of the Drosophila ovary, which correspond to early stages (pachytene) and late stages (metaphase I and beyond) of meiosis. In addition to taking into account morphology, the techniques differ by fixation conditions and the method to isolate the tissue. Most of these methods are whole mounts, which preserve the three-dimensional structure.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. King, R. C. (1970) Ovarian development in Drosophila melanogaster, Academic Press, Inc., New York.

    Google Scholar 

  2. Mahowald, A. P. and Kambysellis, M. P. (1980) Oogenesis, in The Genetics and Biology of Drosophila, Vol 2 d (Ashburner, M. and Wright, T. R. F., eds.), Academic Press, New York, pp. 141–224.

    Google Scholar 

  3. Spradling, A. C. (1993) Developmental genetics of oogenesis, in The Development of Drosophila melanogaster (Bate, M. and Arias, A. M., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 1–70.

    Google Scholar 

  4. Carpenter, A. T. C. (1979) Synaptonemal complex and recombination nodules in wild-type Drosophila melanogaster females. Genetics. 92, 511–541.

    PubMed  CAS  Google Scholar 

  5. Mehrotra, S. and McKim, K. S. (2006) Temporal analysis of meiotic DNA double-strand break formation and repair in Drosophila females. PLoS Genet. 2, e200.

    Article  PubMed  CAS  Google Scholar 

  6. de Cuevas, M., Lilly, M. A., and Spradling, A. C. (1997) Germline cyst formation in Drosophila. Ann. Rev. Genet. 31, 405–428.

    Article  Google Scholar 

  7. Riechmann, V. and Ephrussi, A. (2001) Axis formation during Drosophilaoogenesis. Curr. Opin. Genet. Dev. 11, 374–383.

    Article  CAS  Google Scholar 

  8. Ashburner, M., Golic, K. G., and Hawley, R. S. (2005) Drosophila: a laboratory handbook, 2 ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  9. Sullivan, W., Ashburner, M., and Hawley, R. S. (2000) Drosophila Protocols, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  10. Giunta, K. L., Jang, J. K., Manheim, E. M., Subramanian, G., and McKim, K. S. (2002) subitoencodes a kinesin-like protein required for meiotic spindle pole formation in Drosophila melanogaster. Genetics. 160, 1489–1501.

    PubMed  CAS  Google Scholar 

  11. Belmont, A. S., Braunfeld, M. B., Sedat, J. W., and Agard, D. A. (1989) Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro. Chromosoma. 98, 129–143.

    Article  PubMed  CAS  Google Scholar 

  12. Webber, H. A., Howard, L., and Bickel, S. E. (2004) The cohesion protein ORD is required for homologue bias during meiotic recombination. J. Cell Biol. 164, 819–829.

    Article  PubMed  CAS  Google Scholar 

  13. Dernburg, A. F., Sedat, J. W., and Hawley, R. S. (1996) Direct evidence of a role for heterochromatin in meiotic chromosome segregation. Cell. 85, 135–146.

    Article  Google Scholar 

  14. Sherizen, D., Jang, J. K., Bhagat, R., Kato, N., and McKim, K. S. (2005) Meiotic recombination in Drosophila females depends on chromosome continuity between genetically defined boundaries. Genetics. 169, 767–781.

    Article  PubMed  CAS  Google Scholar 

  15. Gong, W. J., McKim, K. S., and Hawley, R. S. (2005) All paired up with no place to go: Pairing, synapsis, and DSB formation in a balancer heterozygote. PLoS Genet. 1, e67.

    Article  PubMed  Google Scholar 

  16. Theurkauf, W. E. and Hawley, R. S. (1992) Meiotic spindle assembly in Drosophila females: behavior of nonexchange chromosomes and the effects of mutations in the nod kinesin-like protein. J. Cell Biol. 116, 1167–1180.

    Article  PubMed  CAS  Google Scholar 

  17. Mahowald, A. P., Goralski, T. J., and Caulton, J. H. (1983) In vitro activation of Drosophila eggs. Dev. Biol. 98, 437–445.

    Article  PubMed  CAS  Google Scholar 

  18. Endow, S. A. and Komma, D. J. (1997) Spindle dynamics during meiosis in Drosophila oocytes. J. Cell Biol. 137, 1321–1336.

    Article  PubMed  CAS  Google Scholar 

  19. Skold, H. N., Komma, D. J., and Endow, S. A. (2005) Assembly pathway of the anastral Drosophila oocyte meiosis I spindle. J. Cell Sci. 118, 1745–1755.

    Article  PubMed  Google Scholar 

  20. Grieder, N. C., de Cuevas, M., and Spradling, A. C. (2000) The fusome organizes the microtubule network during oocyte differentiation in Drosophila. Development. 127, 4253–4264.

    PubMed  CAS  Google Scholar 

  21. Clarkson, M. and Saint, R. (1999) A His2AvDGFPfusion gene complements a lethal His2AvDmutant allele and provides an in vivomarker for Drosophilachromosome behavior. DNA Cell Biol. 18, 457–462.

    Article  PubMed  CAS  Google Scholar 

  22. Matthies, H. J., McDonald, H. B., Goldstein, L. S., and Theurkauf, W. E. (1996) Anastral meiotic spindle morphogenesis: role of the non-claret disjunctional kinesin-like protein. J. Cell Biol. 134, 455–464.

    Article  PubMed  CAS  Google Scholar 

  23. Matthies, H. J., Clarkson, M., Saint, R. B., Namba, R., and Hawley, R. S. (2000) Analysis of meiosis in fixed and live oocytes by light microscopy, in Drosophila Protocols (Sullivan, W., Ashburner, M., and Hawley, R. S., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 67–85.

    Google Scholar 

  24. Endow, S. A. and Komma, D. J. (1998) Assembly and dynamics of an anastral:astral spindle: the meiosis II spindle of Drosophila oocytes. J. Cell Sci. 111, 2487–2495.

    PubMed  CAS  Google Scholar 

  25. Heifetz, Y., Yu, J., and Wolfner, M. F. (2001) Ovulation triggers activation of Drosophila oocytes. Dev. Biol. 234, 416–424.

    Article  PubMed  CAS  Google Scholar 

  26. Puro, J. and Nokkala, S. (1977) Meiotic segregation of chromosomes in Drosophila melanogaster oocytes. Chromosoma. 63, 273–286.

    Article  Google Scholar 

  27. Williams, B. C., Dernburg, A. F., Puro, J., Nokkala, S., and Goldberg, M. L. (1997) The Drosophilakinesin-like protein KLP3A is required for proper behavior of male and female pronucleii at fertilization. Development. 124, 2365–2376.

    PubMed  CAS  Google Scholar 

  28. Rothwell, W. F. and Sullivan, W. (2000) Fluorescent analysis of Drosophila embryos, in Drosophila Protocols (Sullivan, W., Ashburner, M., and Hawley, R. S., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 141–157.

    Google Scholar 

  29. Gonzalez, C. and Glover, D. M. (1993) Techniques for studying mitosis in Drosophila, in The Cell Cycle (Fantes, P. and Brookes, R., eds.), Oxford University Press, Oxford, UK, pp. 163–168.

    Google Scholar 

  30. Riparbelli, M. G., Callaini, G., Glover, D. M., and Avides, M. C. (2002) A requirement for the Abnormal Spindle protein to organise microtubules of the central spindle for cytokinesis in Drosophila. J. Cell Sci. 115, 913–922.

    PubMed  CAS  Google Scholar 

  31. Page, A. W. and Orr-Weaver, T. L. (1997) Activation of the meiotic divisions in Drosophila oocytes. Dev. Biol. 183, 195–207.

    Article  PubMed  CAS  Google Scholar 

  32. Robb, J. A. (1969) Maintenance of imaginal discs of Drosophila melanogaster in chemically defined media. J. Cell Biol. 41, 876–885.

    Article  PubMed  CAS  Google Scholar 

  33. Page, S. L. and Hawley, R. S. (2001) c(3)G encodes a Drosophila synaptonemal complex protein. Genes Dev. 15, 3130–3143.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to the many Drosophila researchers who have contributed to the development of these methods, including Sharon Bickel, Abby Dernburg, Scott Hawley, Terry Orr-Weaver, Rob Saint, and William Thuerkauf. This work was supported by grants from the NSF (MCB-0719010) and NIH (R01GM067142).

Author information

Authors and Affiliations

  1. Waksman Institute and Department of Genetics, Rutgers, the State University of New Jersey, Piscataway, NJ, USA

    Kim S. McKim, Eric F. Joyce & Janet K. Jang

Authors
  1. Kim S. McKim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eric F. Joyce
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Janet K. Jang
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, York Ave. 1275, New York, 10021-6007, U.S.A.

    Scott Keeney

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

McKim, K.S., Joyce, E.F., Jang, J.K. (2009). Cytological Analysis of Meiosis in Fixed Drosophila Ovaries. 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_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-103-5_12

  • 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

Publish with us

Policies and ethics

Search

Navigation