Advertisement

A Practical Guide to Live-Cell Imaging of Meiosis in Arabidopsis

  • Maria Ada Prusicki
  • Yuki Hamamura
  • Arp SchnittgerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)

Abstract

Plants are powerful model systems to study meiosis. Our knowledge about the cytology of plant meiosis is mainly based on the analysis of fixed material. Although highly informative, this approach is limited in understanding the dynamics of meiosis. Here, we present a step-by-step instruction for a newly developed method to follow meiosis in male meiocytes of Arabidopsis in real time by confocal laser scanning microscopy. We envision that this method can be easily translated to other plant species and especially crops (e.g., Brassica, maize, and potato).

Key words

Meiosis Arabidopsis Confocal laser scanning microscopy Live cell imaging Time lapse 

Notes

Acknowledgments

This work was supported by the EU-ITN grant COMREC, the BMBF grant TarBreed, and core funding of the University of Hamburg to A.S.

References

  1. 1.
    Klutstein M, Fennell A, Fernández-Álvarez A et al (2015) The telomere bouquet regulates meiotic centromere assembly. Nat Cell Biol 17:458–469CrossRefGoogle Scholar
  2. 2.
    Koszul R, Kleckner N (2009) Dynamic chromosome movements during meiosis: a way to eliminate unwanted connections? Trends Cell Biol 19:716–724CrossRefGoogle Scholar
  3. 3.
    Mullen TJ, Wignall SM (2017) Interplay between microtubule bundling and sorting factors ensures acentriolar spindle stability during C. elegans oocyte meiosis. PLoS Genet 13:e1006986CrossRefGoogle Scholar
  4. 4.
    Rosu S, Cohen-Fix O (2017) Live-imaging analysis of germ cell proliferation in the C. elegans adult supports a stochastic model for stem cell proliferation. Dev Biol 423:93–100CrossRefGoogle Scholar
  5. 5.
    Mlynarczyk-Evans S, Villeneuve AM (2017) Time-course analysis of early meiotic prophase events informs mechanisms of homolog pairing and synapsis in Caenorhabditis elegans. Genetics 207:103–114CrossRefGoogle Scholar
  6. 6.
    Enguita-Marruedo A, Cappellen WAV, Hoogerbrugge JW et al (2018) Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries. Chromosoma 127(3):341–359CrossRefGoogle Scholar
  7. 7.
    Holubcová Z, Howard G, Schuh M (2013) Vesicles modulate an actin network for asymmetric spindle positioning. Nat Cell Biol 15:937CrossRefGoogle Scholar
  8. 8.
    Clift D, Schuh M (2013) Restarting life: fertilization and the transition from meiosis to mitosis. Nat Rev Mol Cell Biol 14:549CrossRefGoogle Scholar
  9. 9.
    Kyogoku H, Kitajima TS (2017) Large cytoplasm is linked to the error-prone nature of oocytes. Dev Cell 41:287–298.e4CrossRefGoogle Scholar
  10. 10.
    Yu H-G, Hiatt EN, Chan A et al (1997) Neocentromere-mediated chromosome movement in maize. J Cell Biol 139:831–840CrossRefGoogle Scholar
  11. 11.
    Sheehan MJ, Pawlowski WP (2009) Live imaging of rapid chromosome movements in meiotic prophase I in maize. PNAS 106:20989–20994CrossRefGoogle Scholar
  12. 12.
    Nannas NJ, Higgins DM, Dawe RK (2016) Anaphase asymmetry and dynamic repositioning of the division plane during maize meiosis. J Cell Sci 129:4014–4024PubMedGoogle Scholar
  13. 13.
    Ingouff M, Selles B, Michaud C et al (2017) Live-cell analysis of DNA methylation during sexual reproduction in Arabidopsis reveals context and sex-specific dynamics controlled by noncanonical RdDM. Genes Dev 31:72–83CrossRefGoogle Scholar
  14. 14.
    Wijnker E, Schnittger A (2013) Control of the meiotic cell division program in plants. Plant Reprod 26:143–158CrossRefGoogle Scholar
  15. 15.
    Prusicki MA, Keizer EM, Rosmalen RP van et al (2019) Live cell imaging of meiosis in Arabidopsis thaliana—a landmark system. eLife. 8:e42834.  https://doi.org/10.7554/eLife.42834
  16. 16.
    Yang C, Hamamura Y, Sofroni K et al (2019) SWITCH 1/DYAD is a novel WINGS APART-LIKE antagonist that maintains sister chromatid cohesion in meiosis. Nat Commun. 10:1755.  https://doi.org/10.1038/s41467-019-09759-w
  17. 17.
    Hamant O, Das P, Burian A (2014) Time-lapse imaging of developing meristems using confocal laser scanning microscope. In: Plant cell morphogenesis. Humana Press, Totowa, NJ, pp 111–119CrossRefGoogle Scholar
  18. 18.
    Armstrong SJ, Franklin FCH, Jones GH (2003) A meiotic time-course for Arabidopsis thaliana. Sex Plant Reprod 16:141–149CrossRefGoogle Scholar
  19. 19.
    Stronghill PE, Azimi W, Hasenkampf CA (2014) A novel method to follow meiotic progression in Arabidopsis using confocal microscopy and 5-ethynyl-2′-deoxyuridine labeling. Plant Methods 10:33CrossRefGoogle Scholar
  20. 20.
    Sanchez-Moran E, Santos J-L, Jones GH et al (2007) ASY1 mediates AtDMC1-dependent interhomolog recombination during meiosis in Arabidopsis. Genes Dev 21:2220–2233CrossRefGoogle Scholar
  21. 21.
    Cai X, Dong F, Edelmann RE et al (2003) The Arabidopsis SYN1 cohesin protein is required for sister chromatid arm cohesion and homologous chromosome pairing. J Cell Sci 116:2999–3007CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Maria Ada Prusicki
    • 1
  • Yuki Hamamura
    • 1
  • Arp Schnittger
    • 1
    Email author
  1. 1.Department of Developmental BiologyUniversity of HamburgHamburgGermany

Personalised recommendations