Letter | Published:

Chiral blastomere arrangement dictates zygotic left–right asymmetry pathway in snails

Nature volume 462, pages 790794 (10 December 2009) | Download Citation

Abstract

Most animals display internal and/or external left–right asymmetry. Several mechanisms for left–right asymmetry determination have been proposed for vertebrates1,2,3,4,5,6,7,8,9,10 and invertebrates1,2,4,9,11,12,13,14 but they are still not well characterized, particularly at the early developmental stage. The gastropods Lymnaea stagnalis and the closely related Lymnaea peregra have both the sinistral (recessive) and the dextral (dominant) snails within a species and the chirality is hereditary, determined by a single locus that functions maternally15,16,17,18. Intriguingly, the handedness-determining gene(s) and the mechanisms are not yet identified. Here we show that in L. stagnalis, the chiral blastomere arrangement at the eight-cell stage (but not the two- or four-cell stage) determines the left–right asymmetry throughout the developmental programme, and acts upstream of the Nodal signalling pathway. Thus, we could demonstrate that mechanical micromanipulation of the third cleavage chirality (from the four- to the eight-cell stage) leads to reversal of embryonic handedness. These manipulated embryos grew to ‘dextralized’ sinistral and ‘sinistralized’ dextral snails—that is, normal healthy fertile organisms with all the usual left–right asymmetries reversed to that encoded by the mothers’ genetic information. Moreover, manipulation reversed the embryonic nodal expression patterns. Using backcrossed F7 congenic animals, we could demonstrate a strong genetic linkage between the handedness-determining gene(s) and the chiral cytoskeletal dynamics at the third cleavage that promotes the dominant-type blastomere arrangement. These results establish the crucial importance of the maternally determined blastomere arrangement at the eight-cell stage in dictating zygotic signalling pathways in the organismal chiromorphogenesis. Similar chiral blastomere configuration mechanisms may also operate upstream of the Nodal pathway in left–right patterning of deuterostomes/vertebrates.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Data deposits

Sequences of L. stagnalis nodal and Pitx are deposited at GenBank, with accession numbers respectively GU073383 and GU073384.

References

  1. 1.

    & The development of handedness in left/right asymmetry. Development 109, 1–9 (1990)

  2. 2.

    , , & Strategies to establish left/right asymmetry in vertebrates and invertebrates. Curr. Opin. Genet. Dev. 17, 351–358 (2007)

  3. 3.

    & The left-right axis in the mouse: from origin to morphology. Development 133, 2095–2104 (2006)

  4. 4.

    & Perspectives and open problems in the early phases of left-right patterning. Semin. Cell Dev. Biol. 20, 456–463 (2009)

  5. 5.

    , , & Determination of left-right patterning of the mouse embryo by artificial nodal flow. Nature 418, 96–99 (2002)

  6. 6.

    , , , & Mechanism of nodal flow: a conserved symmetry breaking event in left-right axis determination. Cell 121, 633–644 (2005)

  7. 7.

    et al. De novo formation of left-right asymmetry by posterior tilt of nodal cilia. PLoS Biol. 3, e268 (2005)

  8. 8.

    , , & Nodal flow and the generation of left-right asymmetry. Cell 125, 33–45 (2006)

  9. 9.

    & Left-right patterning from the inside out: widespread evidence for intracellular control. Bioessays 29, 271–287 (2007)

  10. 10.

    , , , & Asymmetries in H+/K+-ATPase and cell membrane potentials comprise a very early step in left-right patterning. Cell 111, 77–89 (2002)

  11. 11.

    , & Type ID unconventional myosin controls left-right asymmetry in Drosophila. Nature 440, 803–807 (2006)

  12. 12.

    et al. An unconventional myosin in Drosophila reverses the default handedness in visceral organs. Nature 440, 798–802 (2006)

  13. 13.

    Evidence from reversal of handedness in C. elegans embryos for early cell interactions determining cell fates. Nature 349, 536–538 (1991)

  14. 14.

    et al. Embryonic handedness choice in C. elegans involves the Gα protein GPA-16. Development 130, 5731–5740 (2003)

  15. 15.

    , , , & The inheritance of sinistrality in Lymnaea peregra. Phil. Trans. R. Soc. Lond. B 219, 51–131 (1930)

  16. 16.

    Inheritance of direction of coiling in Lymnaea. Science 58, 269–270 (1923)

  17. 17.

    & The developmental genetics of dextrality and sinistrality in the gastropod Lymnaea peregra. Wilhelm Roux Arch. Dev. Biol. 191, 69–83 (1982)

  18. 18.

    , & Construction of a backcross progeny collection of dextral and sinistral individuals of a freshwater gastropod, Lymnaea stagnalis. Dev. Genes Evol. 213, 193–198 (2003)

  19. 19.

    Reversal of cleavage in a sinistral gastropod. Ann. NY Acad. Sci. 8, 167–170 (1894)

  20. 20.

    , & Body handedness is directed by genetically determined cytoskeletal dynamics in the early embryo. Curr. Biol. 14, 1462–1467 (2004)

  21. 21.

    & Asymmetrical rotations of blastomeres in early cleavage of gastropoda. Wilhelm Roux Arch. Dev. Biol. 177, 193–203 (1975)

  22. 22.

    & Left-right asymmetry: more than one way to coil a shell. Curr. Biol. 14, R654–R656 (2004)

  23. 23.

    & Evolutionary implications of the mode of D quadrant specification in coelomates with spiral cleavage. J. Evol. Biol. 5, 205–247 (2002)

  24. 24.

    & A conserved role for the nodal signaling pathway in the establishment of dorso-ventral and left-right axes in deuterostomes. J. Exp. Zool. B 310, 41–53 (2008)

  25. 25.

    & Nodal signalling is involved in left-right asymmetry in snails. Nature 457, 1007–1011 (2009)

  26. 26.

    , & Expression of Patella vulgata orthologs of engrailed and dpp-BMP2/4 in adjacent domains during molluscan shell development suggests a conserved compartment boundary mechanism. Dev. Biol. 246, 341–355 (2002)

  27. 27.

    et al. Pitx genes in Tunicates provide new molecular insight into the evolutionary origin of pituitary. Gene 287, 107–113 (2002)

Download references

Acknowledgements

We thank G. Smit for his gift of dextral and sinistral stock of L. stagnalis. We also thank K. Miyoshi, Y. Ozawa and H. Kuwata of the Kuroda Chiromorphology team for their help in rearing snails and creating F7 congenic snails. K. Fujikura, A. Okubo and G. Sai are thanked for their preliminary attempts at in situ hybridization experiments.

Author Contributions R.K. conceived the study, designed/coordinated the experiments and wrote the manuscript. B.E. performed the reversal experiments and whole mount in situ hybridization (WISH) on reversed embryos. M.A. performed WISH on control and F7 congenic snails. M.S. cloned and characterized nodal and Pitx from L. stagnalis to make template vector for the WISH probes. B.E. and M.S. provided comments on the manuscript.

Author information

Affiliations

  1. Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo 153-8902, Japan

    • Reiko Kuroda
  2. Kuroda Chiromorphology Team, ERATO-SORST, JST, Komaba, Meguro-ku, Tokyo 153-0041, Japan

    • Reiko Kuroda
    • , Bunshiro Endo
    • , Masanori Abe
    •  & Miho Shimizu
  3. Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

    • Reiko Kuroda

Authors

  1. Search for Reiko Kuroda in:

  2. Search for Bunshiro Endo in:

  3. Search for Masanori Abe in:

  4. Search for Miho Shimizu in:

Corresponding author

Correspondence to Reiko Kuroda.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-2 with Legends and Supplementary Table 1.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature08597

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.