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Methods Cell Biol. 2018;145:237-248. doi: 10.1016/bs.mcb.2018.03.026. Epub 2018 Apr 11.

Combining microscopy and biochemistry to study meiotic spindle assembly in Drosophila oocytes.

Author information

1
Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
2
Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom. Electronic address: h.ohkura@ed.ac.uk.

Abstract

Studies using Drosophila have played pivotal roles in advancing our understanding of molecular mechanisms of mitosis throughout the past decades, due to the short generation time and advanced genetic research of this organism. Drosophila is also an excellent model to study female meiosis in oocytes. Pathways such as the acentrosomal assembly of the meiotic spindle in oocytes are conserved from fly to humans. Collecting and manipulating large Drosophila oocytes for microscopy and biochemistry are both time and cost efficient, offering advantages over mouse or human oocytes. Therefore, Drosophila oocytes serve as an excellent platform for molecular studies of female meiosis using a combination of genetics, microscopy, and biochemistry. Here we describe key methods to observe the formation of the meiotic spindle either in fixed or in live oocytes. Moreover, biochemical methods are described to identify protein-protein interactions in vivo.

KEYWORDS:

Biochemistry; Drosophila; Fluorescent in situ hybridization; Immunoprecipitation; Immunostaining; Live imaging; Microtubule; Oocyte; Protein–protein interaction; RNAi; Spindle; Transgenesis

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