Article

Anatomical integration of the sacral–hindlimb unit coordinated by GDF11 underlies variation in hindlimb positioning in tetrapods

  • Nature Ecology & Evolution 113921399 (2017)
  • doi:10.1038/s41559-017-0247-y
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Abstract

Elucidating how body parts from different primordia are integrated during development is essential for understanding the nature of morphological evolution. In tetrapod evolution, while the position of the hindlimb has diversified along with the vertebral formula, the mechanism responsible for this coordination has not been well understood. However, this synchronization suggests the presence of an evolutionarily conserved developmental mechanism that coordinates the positioning of the hindlimb skeleton derived from the lateral plate mesoderm with that of the sacral vertebrae derived from the somites. Here we show that GDF11 secreted from the posterior axial mesoderm is a key factor in the integration of sacral vertebrae and hindlimb positioning by inducing Hox gene expression in two different primordia. Manipulating the onset of GDF11 activity altered the position of the hindlimb in chicken embryos, indicating that the onset of Gdf11 expression is responsible for the coordinated positioning of the sacral vertebrae and hindlimbs. Through comparative analysis with other vertebrate embryos, we also show that each tetrapod species has a unique onset timing of Gdf11 expression, which is tightly correlated with the anteroposterior levels of the hindlimb bud. We conclude that the evolutionary diversity of hindlimb positioning resulted from heterochronic shifts in Gdf11 expression, which led to coordinated shifts in the sacral–hindlimb unit along the anteroposterior axis.

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Acknowledgements

We thank Y. Yamamoto-Shiraishi for discussions; G. Sheng for help with collecting the emu embryos; and A. Sakai for help with collecting the snake embryos. This work was supported by JST PRESTO, Grants-in-Aid for Scientific Research—KAKENHI grant numbers 2529150, 15KT0150, 16H01444 and 17H05764.

Author information

Author notes

  1. Atsushi Kuroiwa and Takayuki Suzuki jointly supervised this work.

Affiliations

  1. Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, 464-8602, Japan

    • Yoshiyuki Matsubara
    • , Takaya Suganuma
    • , Yuhei Kohara
    • , Tatsuya Nagai
    • , Atsushi Kuroiwa
    •  & Takayuki Suzuki
  2. Evolutionary Morphology Laboratory, Kobe, 650-0047, Japan

    • Tatsuya Hirasawa
    •  & Shigeru Kuratani
  3. Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, Aoba-ku, Sendai, 980-8578, Japan

    • Shiro Egawa
    •  & Koji Tamura
  4. Institute of Development, Aging and Cancer, Tohoku University, Aoba-ku Sendai, 980-8575, Japan

    • Ayumi Hattori

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Contributions

Y.M., A.K. and T.Suz. conceived the project and designed the experiments. Y.M. performed most experiments; T.H. performed the statistical analyses; S.E. collected ocelot gecko embryos; A.H. performed implantation experiments. T.Sug. and Y.K. performed the cell-tracking study for African clawed frog embryos; T.N. performed gene-expression studies; K.T. contributed ocelot gecko embryos; and Y.M., T.H., S.K., A.K and T.Suz. wrote the paper, with comments from co-authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Atsushi Kuroiwa or Takayuki Suzuki.

Electronic supplementary material

  1. 1.

    Supplementary information

    Supplementary Texts 1–6; Supplementary Figures 1–9; Supplementary References

  2. 2.

    Supplementary Video

    Animated model regarding anatomical integration of the sacral–hindlimb unit and the mechanism that brought about variation in hindlimb positioning in tetrapod evolution.