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Dishevelled controls apical docking and planar polarization of basal bodies in ciliated epithelial cells

Abstract

The planar cell polarity (PCP) signaling system governs many aspects of polarized cell behavior. Here, we use an in vivo model of vertebrate mucociliary epithelial development to show that Dishevelled (Dvl) is essential for the apical positioning of basal bodies. We find that Dvl and Inturned mediate the activation of the Rho GTPase specifically at basal bodies, and that these three proteins together mediate the docking of basal bodies to the apical plasma membrane. Moreover, we find that this docking involves a Dvl-dependent association of basal bodies with membrane-bound vesicles and the vesicle-trafficking protein, Sec8. Once docked, basal bodies again require Dvl and Rho for the planar polarization that underlies directional beating of cilia. These results demonstrate previously undescribed functions for PCP signaling components and suggest that a common signaling apparatus governs both apical docking and planar polarization of basal bodies.

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Figure 1: Dvl is essential for ciliogenesis in multi-ciliated cells.
Figure 2: Dvl is essential for normal cytoskeletal organization in multi-ciliated cells.
Figure 3: Dvl and Inturned are essential for the apical positioning of basal bodies.
Figure 4: Dvl localizes near the base of cilia in multi-ciliated cells.
Figure 5: Dvl and Inturned control the activation and localization of the Rho GTPase.
Figure 6: Dvl links basal body docking to vesicle traffic and Sec8 localization.
Figure 7: Dvl governs planar polarization of basal bodies.
Figure 8: Dvl and Rho govern directional fluid flow across the ciliated epidermis.

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Acknowledgements

We thank R. Gray, J. Hayes and E. Herrington for technical assistance, The Institute for Cellular and Molecular Biology at University of Texas, Austin Core Facility for help with electron microscopy and J. Kirk with Zeiss Microimaging for help with high-speed confocal imaging. We thank A. Ewald for critical reading and discussion. T.J.P. is supported by the William S. Livingston Graduate Fellowship from University of Texas, Austin. This work was supported by US National Institutes of Health/National Institute of General Medical Sciences grants to J.B.W. and C.K. and by grants to J.B.W. from The Sandler Program for Asthma Research, The Burroughs-Wellcome Fund and The March of Dimes.

Author information

Authors and Affiliations

Authors

Contributions

T.J.P., B.J.M., C.K. and J.B.W. designed the study; T.J.P., B.J.M. and P.B.A. carried out the experiments; all authors contributed to the interpretation of the results and the writing of the paper.

Corresponding author

Correspondence to John B Wallingford.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10 (PDF 15678 kb)

Supplementary Movie 1

Flow of beads across a control tadpole is highly organized. (MOV 2770 kb)

Supplementary Movie 2

Flow of beads across a Rho-N19 expressing tadpole is less organized. Swirling and turning of beads can be observed frequently. (MOV 3383 kb)

Supplementary Movie 3

Flow of beads across an Xdd1 expressing tadpole is less organized. Swirling and turning of beads can be observed frequently. (MOV 1500 kb)

Supplementary Movie 4

High-speed confocal time-lapse of beating cilia from an intact control embryo labelled with tau-GFP. (MOV 2483 kb)

Supplementary Movie 5

High-speed confocal time-lapse of beating cilia from an intact embryo labelled with tau-GFP and expressing Xdd1. Beat frequency is similar, but beating appears disorganized. (MOV 1784 kb)

Supplementary Movie 6

High-speed confocal time-lapse of multiple ciliated cells from a control embryo. (MOV 2545 kb)

Supplementary Movie 7

High-speed confocal time-lapse of multiple ciliated cells from an Xdd1 expressing embryo. Overall beat frequency is similar to control, but appears modestly reduced in cells on the left. Cells on the right appear to have a disorganized beating pattern. (MOV 1918 kb)

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Park, T., Mitchell, B., Abitua, P. et al. Dishevelled controls apical docking and planar polarization of basal bodies in ciliated epithelial cells. Nat Genet 40, 871–879 (2008). https://doi.org/10.1038/ng.104

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