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Analysis of Chromosome Dynamics and Chromosomal Proteins in Drosophila Spermatocytes

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Meiosis

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

Abstract

A wide variety of techniques have been utilized to determine the localization of various proteins from premeiotic through meiotic stages in Drosophila males. Live imaging has been instrumental in monitoring chromosome pairing and the localization of fusion proteins. Immunofluorescence has been a widely utilized technique to examine the localization and colocalization of the many proteins involved in meiosis. Recently, an immuno-FISH protocol was developed to observe the co-localization of DNA probes and proteins. In this chapter, detailed protocols outlining these three types of experiments are presented.

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References

  1. Hardy, R. W., Tokuyasu, K. T., Lindsley, D. L., and Garavito, M. (1979) The germinal proliferation center in the testis of Drosophila melanogaster. J. Ultrastruct. Res. 69, 180–90.

    Article  PubMed  CAS  Google Scholar 

  2. Lindsley, D. L. and Tokuyasu., K. T. (1980) Spermatogenesis, in “The genetics and biology of Drosophila” (Ashburner, M. and Wright, T. R. F., Eds.), Vol. 2b, pp. 225–94, Academic Press, London.

    Google Scholar 

  3. Fuller, M. (1993) Spermatogenesis, in “Development of Drosophila” (Martinez-Arias, A. and Bate, M., Eds.), pp. 71–147, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  4. Bonaccorsi, S., Giansanti, M. G., Cenci, G., and Gatti, M. (2000) Cytological analysis of spermatocyte growth and male meiosis in Drosophila melanogaster, in “Drosophila Protocols” (Sullivan, W., Ashburner, M., and Hawley, R.S., Eds.), pp. 87–109, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  5. Cenci, G., Bonaccorsi, S., Pisano, C., Verni, F., and Gatti, M. (1994) Chromatin and microtubule organization during premeiotic, meiotic and early postmeiotic stages of Drosophila melanogaster spermatogenesis. J. Cell Sci. 107, 3521–34.

    PubMed  CAS  Google Scholar 

  6. Thomas, S. E., Soltani-Bejnood, M., Roth, P., Dorn, R., Logsdon, J. M. Jr., and McKee, B. D. (2005) Identification of two proteins required for conjunction and regular segregation of achiasmate homologs in Drosophila male meiosis. Cell 123, 555–68.

    Article  PubMed  CAS  Google Scholar 

  7. Van Noorden, S. and Polak, J. M. (1983) in “Immunocytochemistry, Practical Applications in Pathology and Biology” (Polak, J. M. and Van. Noorden, S., Eds.), pp. 11–42, Wright PSG, Bristol, England.

    Google Scholar 

  8. Sternberger, L. A. (1979) Immunocytochemistry. Wiley, New York.

    Google Scholar 

  9. Melan, M. A. (1999) in “Immunocytochemical Methods and Protocols” (Jovois, L. C., Ed.), Vol. 115, Humana Press, Totowa, NJ.

    Google Scholar 

  10. Gunsalus, K. C., Bonaccorsi, S., Williams, E., Verni, F., Gatti, M., and Goldberg, M. L. (1995) Mutations in twinstar, a Drosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis. J. Cell Biol. 131, 1243–59.

    Article  PubMed  CAS  Google Scholar 

  11. Hime, G. R., Brill, J. A., and Fuller, M. T. (1996) Assembly of ring canals in the male germ line from structural components of the contractile ring. J. Cell Sci. 109, 2779–88.

    PubMed  CAS  Google Scholar 

  12. Gall, J. G. and Pardue, M. L. (1969). Formation and detection of RNA-DNA hybrids in cytological preparations. Proc. Natl. Acad. Sci. U. S. A. 63, 378–83.

    Article  PubMed  CAS  Google Scholar 

  13. Balicky, E. M., Endres, M. W., Lai, C., and Bickel, S. E. (2002) Meiotic cohesion requires accumulation of ORD on chromosomes before condensation. Mol. Biol. Cell 13, 3890–3900.

    Article  PubMed  CAS  Google Scholar 

  14. McKee, B. D. (1996) The license to pair: Identification of meiotic pairing sites in Drosophila. Chromosoma 105, 135–41.

    PubMed  CAS  Google Scholar 

  15. Vazquez, J., Belmont, A. S., and Sedat, J. W. (2002) The dynamics of homologous chromosome pairing during male Drosophila meiosis. Curr. Biol. 12, 1473–83.

    Article  PubMed  CAS  Google Scholar 

  16. Flybase (2008) image gallery. URL: flybase.bio.indiana.edu.

    Google Scholar 

  17. Miller A. (1950) The internal anatomy and histology of the imago of Drosophila melanogaster. in “Biology of Drosophila” (Demerec, M., Ed.), pp. 420–534. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  18. Hartenstein, V. (1993)Atlas of Drosophila Development. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

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Acknowledgements

This work was supported by NIH grant GM40489 to B.D.M.

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Thomas, S.E., McKee, B.D. (2009). Analysis of Chromosome Dynamics and Chromosomal Proteins in Drosophila Spermatocytes. 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_13

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  • DOI: https://doi.org/10.1007/978-1-60761-103-5_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-102-8

  • Online ISBN: 978-1-60761-103-5

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