WNT11 acts as a directional cue to organize the elongation of early muscle fibres

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Abstract

The early vertebrate skeletal muscle is a well-organized tissue in which the primitive muscle fibres, the myocytes, are all parallel and aligned along the antero-posterior axis of the embryo. How myofibres acquire their orientation during development is unknown. Here we show that during early chick myogenesis WNT11 has an essential role in the oriented elongation of the myocytes. We find that the neural tube, known to drive WNT11 expression in the medial border of somites1, is necessary and sufficient to orient myocyte elongation. We then show that the specific inhibition of WNT11 function in somites leads to the disorganization of myocytes. We establish that WNT11 mediates this effect through the evolutionary conserved planar cell polarity (PCP) pathway, downstream of the WNT/β-catenin-dependent pathway, required to initiate the myogenic program of myocytes and WNT11 expression. Finally, we demonstrate that a localized ectopic source of WNT11 can markedly change the orientation of myocytes, indicating that WNT11 acts as a directional cue in this process. All together, these data show that the sequential action of the WNT/PCP and the WNT/β-catenin pathways is necessary for the formation of fully functional embryonic muscle fibres. This study also provides evidence that WNTs can act as instructive cues to regulate the PCP pathway in vertebrates.

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Figure 1: The neural tube is necessary and sufficient for the oriented elongation of the myocytes.
Figure 2: WNT11 regulates the oriented elongation of myocytes through the PCP pathway.
Figure 3: The WNT/β-catenin-dependent pathway, and not the PCP pathway, is required for muscle identity acquisition.
Figure 4: WNT11 acts as an instructive cue during myocyte elongation.

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Acknowledgements

We thank C. Tabin for critical reading of the manuscript. The help of P. Weber for the two-photon imaging, and of the Zeiss team, are acknowledged. We are grateful to M. Manceau for Supplementary Fig. 1d, and to R. Kanadia for his help. This study was funded by grants from the Actions Concertées Incitatives (ACI), the Agence Nationale de la Recherche (ANR), the Association Française contre les Myopathies (AFM) and by the EU 6th Framework Programme Network of Excellence MYORES. J.G. was a Fellow of the AFM.

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Correspondence to Christophe Marcelle.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-5 with `Legends, Supplementary Methods and Supplementary References (PDF 4386 kb)

Supplementary Movie 1

Supplementary Movie 1 shows a 3D reconstruction of a series of 2-photons confocal views (Z-stack) of a somite, 24 hours after electroporation with GFP within the Dorso-Medial Lip (DML). The movie shows the typical bottle-shape morphology of epithelial cells of the DML (in green), of mesenchymal protrusive cells in the transition zone (in blue), and of fully elongated myocytes (in red). (MOV 2254 kb)

Supplementary Movie 2

Supplementary Movie 2 shows a time lapse observation of GFP-electroporated protrusive mesenchymal cells within the transition zone. The movie shows that these cells display an intense protrusive activity characterized by the formation of filopodia extending in all directions and by the generation of lamellipodia at the cell periphery. (MOV 2063 kb)

Supplementary Movie 3

Supplementary Movie 3 shows a time lapse observation of GFP-electroporated cells elongating in the antero-posterior axis of the embryo. The shape of two cells (in red and blue) has been outlined for more clarity. The movie shows that one of them (in red) is elongating in one direction, the second (in blue) in both directions. The formation of full-size myocytes is achieved by an extensive cell elongation driven by the progression of the lamellipodia along the antero-posterior axis of the embryo. (MOV 5462 kb)

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Gros, J., Serralbo, O. & Marcelle, C. WNT11 acts as a directional cue to organize the elongation of early muscle fibres. Nature 457, 589–593 (2009) doi:10.1038/nature07564

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