Live Imaging of Early Mouse Embryos Using Fluorescently Labeled Transgenic Mice

  • Takaya Abe
  • Shinichi Aizawa
  • Toshihiko Fujimori
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1052)

Abstract

Live imaging is a powerful approach to understanding the dynamic processes that occur during development. Periimplantation mouse embryos are transparent, making them suitable for live imaging. In this chapter we describe the culturing and live imaging of mouse embryos, and also introduce the reporter mouse lines for organelle labeling we recently established.

Keywords

Mouse embryo Live imaging Embryo culture Time-lapse observation Reporter mouse lines 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

The authors acknowledge Dr. Kazuki Nakao, Dr. Hiroshi Kiyonari, Dr. Shioi Go, and Mr. Ken-ichi Inoue for producing reporter mouse lines.

References

  1. 1.
    Abe T, Kiyonari H, Shioi G, Inoue K, Nakao K et al (2011) Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging. Genesis 49:579–590PubMedCrossRefGoogle Scholar
  2. 2.
    Nagy A (2003) Manipulating the mouse embryo: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 764 ppGoogle Scholar
  3. 3.
    Hadjantonakis AK, Papaioannou VE (2004) Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice. BMC Biotechnol 4:33PubMedCrossRefGoogle Scholar
  4. 4.
    Kanda T, Sullivan KF, Wahl GM (1998) Histone-GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells. Curr Biol 8:377–385PubMedCrossRefGoogle Scholar
  5. 5.
    Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE et al (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22:1567–1572PubMedCrossRefGoogle Scholar
  6. 6.
    Kurotaki Y, Hatta K, Nakao K, Nabeshima Y, Fujimori T (2007) Blastocyst axis is specified independently of early cell lineage but aligns with the ZP shape. Science 316:719–723PubMedCrossRefGoogle Scholar
  7. 7.
    Rizzuto R, Brini M, Pizzo P, Murgia M, Pozzan T (1995) Chimeric green fluorescent protein as a tool for visualizing subcellular organelles in living cells. Curr Biol 5:635–642PubMedCrossRefGoogle Scholar
  8. 8.
    Llopis J, McCaffery JM, Miyawaki A, Farquhar MG, Tsien RY (1998) Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins. Proc Natl Acad Sci U S A 95:6803–6808PubMedCrossRefGoogle Scholar
  9. 9.
    Teruel MN, Blanpied TA, Shen K, Augustine GJ, Meyer T (1999) A versatile microporation technique for the transfection of cultured CNS neurons. J Neurosci Methods 93:37–48PubMedCrossRefGoogle Scholar
  10. 10.
    Kimble M, Kuzmiak C, McGovern KN, de Hostos EL (2000) Microtubule organization and the effects of GFP-tubulin expression in dictyostelium discoideum. Cell Motil Cytoskeleton 47:48–62PubMedCrossRefGoogle Scholar
  11. 11.
    Faire K, Waterman-Storer CM, Gruber D, Masson D, Salmon ED et al (1999) E-MAP-115 (ensconsin) associates dynamically with microtubules in vivo and is not a physiological modulator of microtubule dynamics. J Cell Sci 112(Pt 23):4243–4255PubMedGoogle Scholar
  12. 12.
    Piehl M, Cassimeris L (2003) Organization and dynamics of growing microtubule plus ends during early mitosis. Mol Biol Cell 14:916–925PubMedCrossRefGoogle Scholar
  13. 13.
    Westphal M, Jungbluth A, Heidecker M, Muhlbauer B, Heizer C et al (1997) Microfilament dynamics during cell movement and chemotaxis monitored using a GFP-actin fusion protein. Curr Biol 7:176–183PubMedCrossRefGoogle Scholar
  14. 14.
    Iioka H, Ueno N, Kinoshita N (2004) Essential role of MARCKS in cortical actin dynamics during gastrulation movements. J Cell Biol 164:169–174PubMedCrossRefGoogle Scholar
  15. 15.
    Nagai T, Ibata K, Park ES, Kubota M, Mikoshiba K et al (2002) A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol 20:87–90PubMedCrossRefGoogle Scholar
  16. 16.
    Petit V, Boyer B, Lentz D, Turner CE, Thiery JP et al (2000) Phosphorylation of tyrosine residues 31 and 118 on paxillin regulates cell migration through an association with CRK in NBT-II cells. J Cell Biol 148:957–970PubMedCrossRefGoogle Scholar
  17. 17.
    Shioi G, Kiyonari H, Abe T, Nakao K, Fujimori T et al (2011) A mouse reporter line to conditionally mark nuclei and cell membranes for in vivo live-imaging. Genesis 49:570–578PubMedCrossRefGoogle Scholar
  18. 18.
    de Felipe P, Luke GA, Hughes LE, Gani D, Halpin C et al (2006) E unum pluribus: multiple proteins from a self-processing polyprotein. Trends Biotechnol 24:68–75PubMedCrossRefGoogle Scholar
  19. 19.
    Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21:70–71PubMedCrossRefGoogle Scholar
  20. 20.
    Srinivas S, Watanabe T, Lin CS, William CM, Tanabe Y et al (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 1:4PubMedCrossRefGoogle Scholar
  21. 21.
    Lakso M, Pichel JG, Gorman JR, Sauer B, Okamoto Y et al (1996) Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. Proc Natl Acad Sci USA 93:5860–5865PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Takaya Abe
    • 1
  • Shinichi Aizawa
    • 1
  • Toshihiko Fujimori
    • 2
  1. 1.Laboratory for Animal Resources and Genetic EngineeringRIKEN Center for Developmental Biology (CDB)KobeJapan
  2. 2.Division of EmbryologyNational Institute for Basic Biology (NIBB)OkazakiJapan

Personalised recommendations