Literature DB >> 29957206

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

Pierre Romé1, Hiroyuki Ohkura2.   

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.
© 2018 Elsevier Inc. All rights reserved.

Entities:  

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

Mesh:

Substances:

Year:  2018        PMID: 29957206      PMCID: PMC6031290          DOI: 10.1016/bs.mcb.2018.03.026

Source DB:  PubMed          Journal:  Methods Cell Biol        ISSN: 0091-679X            Impact factor:   1.441


  34 in total

1.  Msps protein is localized to acentrosomal poles to ensure bipolarity of Drosophila meiotic spindles.

Authors:  C F Cullen; H Ohkura
Journal:  Nat Cell Biol       Date:  2001-07       Impact factor: 28.824

2.  Creating transgenic Drosophila by microinjecting the site-specific phiC31 integrase mRNA and a transgene-containing donor plasmid.

Authors:  Matthew P Fish; Amy C Groth; Michele P Calos; Roel Nusse
Journal:  Nat Protoc       Date:  2007       Impact factor: 13.491

Review 3.  Chromosomal control of meiotic cell division.

Authors:  K S McKim; R S Hawley
Journal:  Science       Date:  1995-12-08       Impact factor: 47.728

4.  Gal4 in the Drosophila female germline.

Authors:  P Rørth
Journal:  Mech Dev       Date:  1998-11       Impact factor: 1.882

Review 5.  Stopping and starting the meiotic cell cycle.

Authors:  A W Page; T L Orr-Weaver
Journal:  Curr Opin Genet Dev       Date:  1997-02       Impact factor: 5.578

6.  The kinesinlike protein Subito contributes to central spindle assembly and organization of the meiotic spindle in Drosophila oocytes.

Authors:  J K Jang; T Rahman; K S McKim
Journal:  Mol Biol Cell       Date:  2005-07-29       Impact factor: 4.138

7.  A genome-scale shRNA resource for transgenic RNAi in Drosophila.

Authors:  Jian-Quan Ni; Rui Zhou; Benjamin Czech; Lu-Ping Liu; Laura Holderbaum; Donghui Yang-Zhou; Hye-Seok Shim; Rong Tao; Dominik Handler; Phillip Karpowicz; Richard Binari; Matthew Booker; Julius Brennecke; Lizabeth A Perkins; Gregory J Hannon; Norbert Perrimon
Journal:  Nat Methods       Date:  2011-04-03       Impact factor: 28.547

8.  Dual roles of Incenp crucial to the assembly of the acentrosomal metaphase spindle in female meiosis.

Authors:  Nathalie Colombié; C Fiona Cullen; Amy L Brittle; Janet K Jang; William C Earnshaw; Mar Carmena; Kim McKim; Hiroyuki Ohkura
Journal:  Development       Date:  2008-08-28       Impact factor: 6.868

9.  The conserved kinase NHK-1 is essential for mitotic progression and unifying acentrosomal meiotic spindles in Drosophila melanogaster.

Authors:  C Fiona Cullen; Amy L Brittle; Takashi Ito; Hiroyuki Ohkura
Journal:  J Cell Biol       Date:  2005-11-21       Impact factor: 10.539

10.  14-3-3 regulation of Ncd reveals a new mechanism for targeting proteins to the spindle in oocytes.

Authors:  Robin Beaven; Ricardo Nunes Bastos; Christos Spanos; Pierre Romé; C Fiona Cullen; Juri Rappsilber; Régis Giet; Gohta Goshima; Hiroyuki Ohkura
Journal:  J Cell Biol       Date:  2017-08-31       Impact factor: 10.539

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