Abstract
Laser ablation is a powerful tool to study forces within biological systems. This technique has been extensively used to study mitotic spindle formation and chromosome segregation. This chapter describes laser ablation of microtubule–chromosome attachments coupled to fluorescence live microscopy and quantitative analysis of individual chromosome movement after microtubule severing. This method allows to gain insight into the organization and dynamics of the meiotic spindle and chromosomes in metaphase I mouse oocytes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bennabi I, Terret M-E, Verlhac M-H (2016) Meiotic spindle assembly and chromosome segregation in oocytes. J Cell Biol 215:611–619
Dumont J, Desai A (2012) Acentrosomal spindle assembly and chromosome segregation during oocyte meiosis. Trends Cell Biol 22:241–249
Bringmann H, Hyman AA (2005) A cytokinesis furrow is positioned by two consecutive signals. Nature 436:731–734
Grill SW, Gönczy P, Stelzer EH et al (2001) Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo. Nature 409:630–633
Riche S, Zouak M, Argoul F et al (2013) Evolutionary comparisons reveal a positional switch for spindle pole oscillations in Caenorhabditis embryos. J Cell Biol 201:653–662
Tolić-Nørrelykke IM, Sacconi L, Thon G et al (2004) Positioning and elongation of the fission yeast spindle by microtubule-based pushing. Curr Biol 14:1181–1186
Brugués J, Nuzzo V, Mazur E et al (2012) Nucleation and transport organize microtubules in metaphase spindles. Cell 149:554–564
Cojoc G, Roscioli E, Zhang L et al (2016) Laser microsurgery reveals conserved viscoelastic behavior of the kinetochore. J Cell Biol 212:767–776
Elting MW, Hueschen CL, Udy DB et al (2014) Force on spindle microtubule minus ends moves chromosomes. J Cell Biol 206:245–256
Kajtez J, Solomatina A, Novak M et al (2016) Overlap microtubules link sister k-fibres and balance the forces on bi-oriented kinetochores. Nat Commun 7:10298
Khodjakov A, La Terra S, Chang F (2004) Laser microsurgery in fission yeast; role of the mitotic spindle midzone in anaphase B. Curr Biol 14:1330–1340
Maiato H, Rieder CL, Khodjakov A (2004) Kinetochore-driven formation of kinetochore fibers contributes to spindle assembly during animal mitosis. J Cell Biol 167:831–840
Sikirzhytski V, Magidson V, Steinman JB et al (2014) Direct kinetochore-spindle pole connections are not required for chromosome segregation. J Cell Biol 206:231–243
Nakagawa S, FitzHarris G (2016) Quantitative microinjection of Morpholino antisense oligonucleotides into mouse oocytes to examine gene function in meiosis-I. Methods Mol Biol Clifton NJ 1457:217–230
Chaigne A, Campillo C, Gov NS et al (2015) A narrow window of cortical tension guides asymmetric spindle positioning in the mouse oocyte. Nat Commun 6:6027
Breuer M, Kolano A, Kwon M et al (2010) HURP permits MTOC sorting for robust meiotic spindle bipolarity, similar to extra centrosome clustering in cancer cells. J Cell Biol 191:1251–1260
Reis A, Chang H-Y, Levasseur M et al (2006) APCcdh1 activity in mouse oocytes prevents entry into the first meiotic division. Nat Cell Biol 8:539–540
Dumont J, Petri S, Pellegrin F et al (2007) A centriole- and RanGTP-independent spindle assembly pathway in meiosis I of vertebrate oocytes. J Cell Biol 176:295–305
Hached K, Xie SZ, Buffin E et al (2011) Mps1 at kinetochores is essential for female mouse meiosis I. Development 138:2261–2271
Schuh M, Ellenberg J (2007) Self-organization of MTOCs replaces centrosome function during acentrosomal spindle assembly in live mouse oocytes. Cell 130:484–498
Kitajima TS, Ohsugi M, Ellenberg J (2011) Complete kinetochore tracking reveals error-prone homologous chromosome biorientation in mammalian oocytes. Cell 146:568–581
de CF, Dallongeville S, Chenouard N et al (2012) Icy: an open bioimage informatics platform for extended reproducible research. Nat Methods 9:690–696
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bennabi, I., Manil-Ségalen, M. (2018). Laser Ablation of Microtubule–Chromosome Attachment in Mouse Oocytes. In: Verlhac, MH., Terret, ME. (eds) Mouse Oocyte Development. Methods in Molecular Biology, vol 1818. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8603-3_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8603-3_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8602-6
Online ISBN: 978-1-4939-8603-3
eBook Packages: Springer Protocols