Abstract
Whole-mount immunofluorescence allows direct visualization of the cellular architecture within cells. Here, we apply this technique to mouse oocytes to visualize spindle morphology and microtubule attachments to kinetochores, using a technique we call “cold treatment,” at various phases of the meiotic cell cycle. This method allows the analysis of spindle structures at different meiosis I stages and at metaphase II. An adaptation of the protocol to the cell cycle stage of interest is described.
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References
Maro B, Verlhac MH (2002) Polar body formation: new rules for asymmetric divisions. Nat Cell Biol 4:E281–E283
Chaigne A, Verlhac MH, Terret ME (2012) Spindle positioning in mammalian oocytes. Exp Cell Res 318:1442–1447
Maro B, Howlett SK, Webb M (1985) Non-spindle microtubule organizing centers in metaphase II-arrested mouse oocytes. J Cell Biol 101:1665–1672
Van Blerkom J (1991) Microtubule mediation of cytoplasmic and nuclear maturation during the early stages of resumed meiosis in cultured mouse oocytes. Proc Natl Acad Sci U S A 88:5031–5035
Schuh M, Ellenberg J (2007) Self-organization of MTOCs replaces centrosome function during acentrosomal spindle assembly in live mouse oocytes. Cell 130:484–498
Clift D, Schuh M (2015) A three-step MTOC fragmentation mechanism facilitates bipolar spindle assembly in mouse oocytes. Nat Commun 6:7217
Courtois A, Yoshida S, Takenouchi O et al (2021) Stable kinetochore-microtubule attachments restrict MTOC position and spindle elongation in oocytes. EMBO Rep 22:e51400
Kitajima TS, Ohsugi M, Ellenberg J (2011) Complete kinetochore tracking reveals error-prone homologous chromosome biorientation in mammalian oocytes. Cell 146:568–581
Schuh M, Ellenberg J (2008) A new model for asymmetric spindle positioning in mouse oocytes. Curr Biol 18:1986–1992
Verlhac MH, Lefebvre C, Guillaud P et al (2000) Asymmetric division in mouse oocytes: with or without Mos. Curr Biol 10:1303–1306
Gryaznova Y, Keating L, Touati SA et al (2021) Kinetochore individualization in meiosis I is required for centromeric cohesin removal in meiosis II. EMBO J 40:e106797
Schatten G, Simerly C, Schatten H (1985) Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. Proc Natl Acad Sci U S A 82:4152–4156
Maro B, Johnson MH, Pickering SJ, Flach G (1984) Changes in actin distribution during fertilization of the mouse egg. J Embryol Exp Morphol 81:211–237
Brinkley BR, Cartwright J (1975) Cold-labile and cold-stable microtubules in the mitotic spindle of mammalian cells. Ann N Y Acad Sci 253:428–439
Touati SA, Buffin E, Cladiere D et al (2015) Mouse oocytes depend on BubR1 for proper chromosome segregation but not for prophase I arrest. Nat Commun 6:6946
Vallot A, Leontiou I, Cladiere D et al (2018) Tension-induced error correction and not kinetochore attachment status activates the SAC in an Aurora-B/C-dependent manner in oocytes. Curr Biol 28:130–139
Acknowledgments
We thank D. Cladière for comments on the manuscript. Research in the group is supported by the Agence Nationale de la Recherche (ANR-19-CE13-0015) and la Fondation de la Recherche Médicale (Equipe FRM DEQ 202103012574).
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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El Jailani, S., Wassmann, K., Touati, S.A. (2024). Whole-Mount Immunofluorescence Staining to Visualize Cell Cycle Progression in Mouse Oocyte Meiosis. In: Castro, A., Lacroix, B. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 2740. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3557-5_13
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DOI: https://doi.org/10.1007/978-1-0716-3557-5_13
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Publisher Name: Humana, New York, NY
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