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Studying Meiosis-Specific Cohesins in Mouse Embryonic Oocytes

  • Ji-hye Kim
  • Kei-ichiro IshiguroEmail author
  • Nobuaki Kudo
  • Yoshinori Watanabe
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 957)

Abstract

Distinct meiotic cohesin complexes play fundamental roles in various meiosis-specific chromosomal events in spatiotemporally different manners during mammalian meiotic prophase. Immunostaining is one of the essential methods to study meiotic cohesin dynamics. For the study of cohesins in the meiotic prophase of oocytes, ovaries should be taken from the embryos during a very limited period before birth. Here we focus on some technical tips concerning the preparation of oocyte chromosome spreads for immunostaining. Further, we describe a method for chromosome fluorescence in situ hybridization (FISH) against immunostained oocytes.

Key words

Fetal oocyte Embryonic ovary Meiosis Cohesin Prophase Axial element Homolog synapsis Synaptonemal complex Chromosome spread FISH 

Notes

Acknowledgements

We thank Dr. Aya Sato for the technical advice on Chromosome FISH with co-immuostaining. This work was supported in part by a JSPS Research Fellowship (to J.K.), a Grant-in-Aid for Scientific Research on Priority Areas, a Grant-in-Aid for Young Scientists (to K.I.), the Global COE Program (Integrative Life Science Based on the Study of Biosignaling Mechanisms), a Grant-in-Aid for Specially Promoted Research (to Y.W.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Ji-hye Kim
    • 1
    • 2
  • Kei-ichiro Ishiguro
    • 1
    Email author
  • Nobuaki Kudo
    • 3
  • Yoshinori Watanabe
    • 1
    • 2
  1. 1.Laboratory of Chromosome Dynamics, Institute of Molecular and Cellular BiosciencesUniversity of TokyoTokyoJapan
  2. 2.Graduate School of Agricultural and Life ScienceUniversity of TokyoTokyoJapan
  3. 3.Institute of Reproductive and Developmental Biology, Imperial College LondonLondonUK

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