Abstract
This chapter describes a technique for performing chromosome spreads from mouse oocytes. It is based on a previously described protocol (Hodges and Hunt, Chromosoma 111: 165–169, 2002), which we have modified. Chromosomes are stained with either Propidium Iodide or Hoechst. This spreading technique allows for simultaneous immunostaining of proteins associated with chromosomes. It is very useful to stain spreads with CREST serum which labels kinetochores, to be able to distinguish bivalents (chromosome pairs), dyads or univalents (paired sister chromatids), and single sister chromatids without ambiguity.
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References
Hodges CA, Hunt PA (2002) Simultaneous analysis of chromosomes and chromosome-associated proteins in mammalian oocytes and embryos. Chromosoma 111:165–169
Nasmyth K, Haering CH (2009) Cohesin: its roles and mechanisms. Annu Rev Genet 43:525–558
Petronczki M, Siomos MF, Nasmyth K (2003) Un menage a quatre: the molecular biology of chromosome segregation in meiosis. Cell 112:423–440
Hassold T, Hunt P (2001) To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev Genet 2:280–291
Revenkova E, Herrmann K, Adelfalk C, Jessberger R (2010) Oocyte cohesin expression restricted to predictyate stages provides full fertility and prevents aneuploidy. Curr Biol 20:1529–1533
Chiang T, Duncan FE, Schindler K, Schultz RM, Lampson MA (2010) Evidence that weakened centromere cohesion is a leading cause of age-related aneuploidy in oocytes. Curr Biol 20:1522–1528
Lister LM, Kouznetsova A, Hyslop LA, Kalleas D, Pace SL, Barel JC, Nathan A, Floros V, Adelfalk C, Watanabe Y, Jessberger R, Kirkwood TB, Hoog C, Herbert M (2010) Age-related meiotic segregation errors in mammalian oocytes are preceded by depletion of cohesin and Sgo2. Curr Biol 20:1511–1521
Hodges CA, Revenkova E, Jessberger R, Hassold TJ, Hunt PA (2005) SMC1beta-deficient female mice provide evidence that cohesins are a missing link in age-related nondisjunction. Nat Genet 37:1351–1355
Daniel K, Lange J, Hached K, Fu J, Anastassiadis K, Roig I, Cooke HJ, Stewart AF, Wassmann K, Jasin M, Keeney S, Toth A (2011) Meiotic homologue alignment and its quality surveillance are controlled by mouse HORMAD1. Nat Cell Biol 13:599–610
Niault T, Hached K, Sotillo R, Sorger PK, Maro B, Benezra R, Wassmann K (2007) Changing Mad2 levels affects chromosome segregation and spindle assembly checkpoint control in female mouse meiosis I. PLoS One 2:e1165
Hached K, Xie SZ, Buffin E, Cladiere D, Rachez C, Sacras M, Sorger PK, Wassmann K (2011) Mps1 at kinetochores is essential for female mouse meiosis I. Development 138:2261–2271
Nicolson GL, Yanagimachi R, Yanagimachi H (1975) Ultrastructural localization of lectin-binding sites on the zonae pellucidae and plasma membranes of mammalian eggs. J Cell Biol 66:263–274
Acknowledgments
We thank S. Touati for comments on the manuscript. K.H. was supported through a PhD fellowship by the Ministère de la Recherche and ARC. Research in the group of K.W. is supported by a grant from La Ligue Nationale Contre Le Cancer (Comité d’Ile de France), the UPMC and CNRS.
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Chambon, JP., Hached, K., Wassmann, K. (2013). Chromosome Spreads with Centromere Staining in Mouse Oocytes. In: Homer, H. (eds) Mammalian Oocyte Regulation. Methods in Molecular Biology, vol 957. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-191-2_14
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DOI: https://doi.org/10.1007/978-1-62703-191-2_14
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