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Live Imaging of X-Chromosome Inactivation and Reactivation Kinetics

  • Shin Kobayashi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)

Abstract

X-chromosome inactivation (XCI) is an epigenetic phenomenon that equalizes the number of X-linked gene products between male and female eutherian mammals by inactivating one of the two X chromosomes. XCI is essential for female mammalian development, and its failure can lead to embryonic death in mutant mice. The pattern of which X chromosome is inactivated changes dynamically during mouse embryogenesis, depending on developmental stages and tissues. Recent progress in molecular biology, including next-generation sequencing (NGS)-based analyses, enables the analysis of gene expression profiles at a single cell level. Combined with NGS technology, live imaging systems can now be used to track epigenetic events and clarify their casual and spatiotemporal relationships to cell differentiation and embryonic development. Here, I describe a novel live-cell imaging system based on “Momiji” mice for monitoring XCI at the single cell level.

Key words

Live imaging X-chromosome inactivation Momiji mouse Preimplantation and postimplantation embryos Single cell analysis Enhanced green fluorescent protein 

Notes

Acknowledgments

I am grateful to Dr. Kazuo Yamagata at Kindai University for technical and experimental support in live imaging of preimplantation mouse embryos, and Dr. Akira Sato at Carl Zeiss Microscopy Co. Ltd. for technical support and discussion on confocal microscopy observations and quantification. This work was supported by the Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology (PRESTO) and by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (Nos. 23500492, 15H01468, and 26430087) to S.K.

References

  1. 1.
    Kobayashi S (2017) Live imaging of X chromosome inactivation and reactivation dynamics. Develop Growth Differ 59:493–500.  https://doi.org/10.1111/dgd.12365CrossRefGoogle Scholar
  2. 2.
    Kobayashi S, Hosoi Y, Shiura H et al (2016) Live imaging of X chromosome reactivation dynamics in early mouse development can discriminate naïve from primed pluripotent stem cells. Development 143:2958–2964.  https://doi.org/10.1242/dev.136739CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Hadjantonakis AK, Cox LL, Tam PP et al (2001) An X-linked GFP transgene reveals unexpected paternal X-chromosome activity in trophoblastic giant cells of the mouse placenta. Genesis 29:133–140CrossRefPubMedCentralGoogle Scholar
  4. 4.
    Takagi N, Sugimoto M, Yamaguchi S et al (2002) Nonrandom X chromosome inactivation in mouse embryos carrying Searle’s T(X;16)16H translocation visualized using X-linked LacZ and GFP transgenes. Cytogenet Genome Res 99:52–58.  https://doi.org/10.1159/000071574CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Takahashi S, Kobayashi S, Hiratani I (2017) Epigenetic differences between naïve and primed pluripotent stem cells. Cell Mol Life Sci.  https://doi.org/10.1007/s00018-017-2703-xCrossRefPubMedCentralGoogle Scholar
  6. 6.
    Nakao K, Nakagata N, Katsuki M (1997) Simple and efficient vitrification procedure for cryopreservation of mouse embryos. Exp Anim 46:231–234CrossRefPubMedCentralGoogle Scholar
  7. 7.
    Lawitts JA, Biggers JD (1993) Culture of preimplantation embryos. Methods Enzymol 225:153–164CrossRefPubMedCentralGoogle Scholar
  8. 8.
    Kimura H, Yamagata K (2015) Visualization of epigenetic modifications in preimplantation embryos. Methods Mol Biol 1222:127–147.  https://doi.org/10.1007/978-1-4939-1594-1_10CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Yamazaki T, Yamagata K, Baba T (2007) Time-lapse and retrospective analysis of DNA methylation in mouse preimplantation embryos by live cell imaging. Dev Biol 304(1):409–419.  https://doi.org/10.1016/j.ydbio.2006.12.046CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Ying QL, Wray J, Nichols J et al (2008) The ground state of embryonic stem cell self-renewal. Nature 453:519–523.  https://doi.org/10.1038/nature06968CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Molecular Profiling Research Center for Drug DiscoveryNational Institute of Advanced Industrial Science and TechnologyTokyoJapan
  2. 2.Department of Epigenetics, Medical Research InstituteTokyo Medical and Dental UniversityTokyoJapan

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