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RhoA and microtubule dynamics control cell–basement membrane interaction in EMT during gastrulation

Nature Cell Biology volume 10, pages 765–775 (2008)Cite this article

An Erratum to this article was published on 01 August 2008

Abstract

Molecular and cellular mechanisms of epithelial–mesenchymal transition (EMT), crucial in development and pathogenesis, are still poorly understood. Here we provide evidence that distinct cellular steps of EMT occur sequentially during gastrulation. Basement membrane (BM) breakdown is the first recognizable step and is controlled by loss of basally localized RhoA activity and its activator neuroepithelial-transforming-protein-1 (Net1). Failure of RhoA downregulation during EMT leads to BM retention and reduction of its activity in normal epithelium leads to BM breakdown. We also show that this is in part mediated by RhoA-regulated basal microtubule stability. Microtubule disruption causes BM breakdown and its stabilization results in BM retention. We propose that loss of Net1 before EMT reduces basal RhoA activity and destabilizes basal microtubules, causing disruption of epithelial cell–BM interaction and subsequently, breakdown of the BM.

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Figure 1: RhoA mis-expression causes EMT defects.
Figure 2: Expression of EMT-related markers.
Figure 3: Effect of RhoA on laminin and distribution of active RhoA.
Figure 4: Net1 expression pattern and its effect on laminin.
Figure 5: Inhibition of RhoA signalling causes premature BM breakdown and rescues mesoderm aggregation.
Figure 6: Effect of nocodazole and taxol on laminin and distribution of microtubules during EMT.
Figure 7: 6G7 recognizes a modified β-tubulin localized to the basal cortex in lateral cells and absent in medial cells.
Figure 8: A model for the regulation of cell–BM interaction by basal RhoA activity and microtubule dynamics.

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Acknowledgements

We thank Fujio Toki for the initial analyses of fibronectin, laminin and 6G7; Yoshiko Takahashi for bringing the phenotype of RhoA cells to our attention; Kathy Joubin for sharing Net1 information; Cantas Alev for generating the Net1 probe; Shigenobu Yonemura and Kazuyo Misaki for help with EM; Masatoshi Takeichi, Yoshiko Takahashi, William Bement, Jeffrey Frost, Alan Hall, Kozo Kaibuchi and Shigenobu Yonemura for sharing reagents; Noriaki Sasai and Yoshiki Sasai for help with Xenopus experiments. Claudio Stern, Donald Newgreen, Kozo Kaibuchi, Masatoshi Takeichi, Fumio Matsuzaki, Shigeo Hayashi and Shigenobu Yonemura for critical comments on the manuscript.

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  1. Laboratory for Early Embryogenesis, RIKEN Center for Developmental Biology, Kobe, 650-0047, Hyogo, Japan

    Yukiko Nakaya, Erike W. Sukowati, Yuping Wu & Guojun Sheng

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  1. Yukiko Nakaya
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Y.N. and G.S. designed the experiments and analysed the data; Y.N, E.W.S. and Y.W. performed the experiments; G.S wrote the manuscript.

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Correspondence to Guojun Sheng.

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Nakaya, Y., Sukowati, E., Wu, Y. et al. RhoA and microtubule dynamics control cell–basement membrane interaction in EMT during gastrulation. Nat Cell Biol 10, 765–775 (2008). https://doi.org/10.1038/ncb1739

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