Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.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.
Figure 1. Dvl is essential for ciliogenesis in multi-ciliated cells. (a) The epidermis of the Xenopus embryo is composed of multi-ciliated cells and mucus-secreting cells. Ciliated cells, labeled by alpha-tubulin immunostaining (red), are evenly spaced between mucus-secreting cells. Polarized beating of cilia generates directional flow across the epithelium16, 26. (b) Normal ciliated cell from a control-injected embryo (injected with a mismatched morpholino). (c) Defective cilia outgrowth in Dvl1–Dvl3 double morphants. (d) Wider field view of control (mismatch morpholino–injected) embryo. (e) Wider field view of Dvl1–Dvl3 double morphant embryo showing many affected cells.
Image redisplayed with permission from Macmillan Publishers Ltd.
Figure 4 - Dvl localizes near the base of cilia in multi-ciliated cells. (a–c) Dvl2 immunostaining shows enrichment at cell membranes throughout the epidermis and enrichment at the apical surface of ciliated cells, as indicated by immunostaining for centrin (b,c). Scale bar, 15 mum. (d) Signal for Dvl2 immunostaining (red) localizes immediately adjacent to the basal body, as indicated by Centrin2-GFP (green). Scale bar, 1 mum. (e) Signal for Dvl2 immunostaining (red) localizes to the center of the region marked by CLAMP-GFP (green); data in Supplementary Figure 4 indicate that CLAMP-GFP labels the ciliary rootlet. Scale bar, 1 mum. (f) The C terminus of Dvl2 is sufficient to drive localization to basal body in multi-ciliated cells. Stage 27 embryos are shown throughout. Scale bar, 5 um.
Image redisplayed with permission from Macmillan Publishers Ltd.
Figure 8 - Dvl and Rho govern directional fluid flow across the ciliated epidermis. (a) Control embryos were grown until stage 26, and latex beads were added to the media to visualize fluid flow over the epidermis. The movement of beads was recorded and representative beads were tracked. Representative bead movements were tracked from the horizontal myoseptum, as indicated by the colored lines. Beads move from dorso-anterior to ventral-posterior uniformly in control embryos. (b) DN-RhoA expression severely disrupts directional flow; beads were turning and swirling frequently. (c) Xdd1 expression severely disrupts directional flow; beads were turning and swirling frequently. (d) The directionality of beads was quantified as the ratio of the linear distance between the first and last point of a track and the total distance along the track. A straight line would be a value of 1.00. Error bars, s.d. (e) DN-RhoA or Xdd1 reduces the rate of bead movement across the epidermis. Defects in polarized flow were evident in that even beads moving in comparatively straight paths moved at a greatly reduced rate across experimental embryos. Error bars, s.d. (f) Still images from a high-speed time-lapse movie of beating cilia labeled with tau-GFP. (g) Still images from a high-speed time-lapse movie of beating cilia expressing Xdd1; frequency is comparable to controls.
Image redisplayed with permission from Macmillan Publishers Ltd.
Adler,
Inturned localizes to the proximal side of wing cells under the instruction of upstream planar polarity proteins.
2004, Pubmed
Adler,
Inturned localizes to the proximal side of wing cells under the instruction of upstream planar polarity proteins.
2004,
Pubmed Ahmad-Annuar,
Signaling across the synapse: a role for Wnt and Dishevelled in presynaptic assembly and neurotransmitter release.
2006,
Pubmed Axelrod,
Differential recruitment of Dishevelled provides signaling specificity in the planar cell polarity and Wingless signaling pathways.
1998,
Pubmed
,
Xenbase Benink,
Concentric zones of active RhoA and Cdc42 around single cell wounds.
2005,
Pubmed
,
Xenbase Bilic,
Wnt induces LRP6 signalosomes and promotes dishevelled-dependent LRP6 phosphorylation.
2007,
Pubmed Boisvieux-Ulrich,
Cytochalasin D inhibits basal body migration and ciliary elongation in quail oviduct epithelium.
1990,
Pubmed Boisvieux-Ulrich,
The orientation of ciliary basal bodies in quail oviduct is related to the ciliary beating cycle commencement.
1985,
Pubmed Capelluto,
The DIX domain targets dishevelled to actin stress fibres and vesicular membranes.
2002,
Pubmed
,
Xenbase Chen,
Dishevelled 2 recruits beta-arrestin 2 to mediate Wnt5A-stimulated endocytosis of Frizzled 4.
2003,
Pubmed Chilvers,
Local mucociliary defence mechanisms.
2000,
Pubmed Classen,
Hexagonal packing of Drosophila wing epithelial cells by the planar cell polarity pathway.
2005,
Pubmed Cohen,
Ciliogenesis and centriole formation in the mouse embryonic nervous system. An ultrastructural analysis.
1988,
Pubmed Dawe,
Centriole/basal body morphogenesis and migration during ciliogenesis in animal cells.
2007,
Pubmed Frisch,
Development of order during ciliogenesis.
1968,
Pubmed Gerdes,
Disruption of the basal body compromises proteasomal function and perturbs intracellular Wnt response.
2007,
Pubmed Gomperts,
Foxj1 regulates basal body anchoring to the cytoskeleton of ciliated pulmonary epithelial cells.
2004,
Pubmed Habas,
Wnt/Frizzled activation of Rho regulates vertebrate gastrulation and requires a novel Formin homology protein Daam1.
2001,
Pubmed
,
Xenbase Hayes,
Identification of novel ciliogenesis factors using a new in vivo model for mucociliary epithelial development.
2007,
Pubmed
,
Xenbase Huang,
Foxj1 is required for apical localization of ezrin in airway epithelial cells.
2003,
Pubmed Jékely,
Evolution of intraflagellar transport from coated vesicles and autogenous origin of the eukaryotic cilium.
2006,
Pubmed Kibar,
Mutations in VANGL1 associated with neural-tube defects.
2007,
Pubmed Kim,
The Bardet-Biedl protein BBS4 targets cargo to the pericentriolar region and is required for microtubule anchoring and cell cycle progression.
2004,
Pubmed Kishida,
Dvl regulates endo- and exocytotic processes through binding to synaptotagmin.
2007,
Pubmed König,
Planar polarity in the ciliated epidermis of Xenopus embryos.
1993,
Pubmed
,
Xenbase Kramer-Zucker,
Cilia-driven fluid flow in the zebrafish pronephros, brain and Kupffer's vesicle is required for normal organogenesis.
2005,
Pubmed Lanzetti,
Actin in membrane trafficking.
2007,
Pubmed Mitchell,
A positive feedback mechanism governs the polarity and motion of motile cilia.
2007,
Pubmed
,
Xenbase Nachury,
A core complex of BBS proteins cooperates with the GTPase Rab8 to promote ciliary membrane biogenesis.
2007,
Pubmed Oishi,
Regulation of primary cilia formation and left-right patterning in zebrafish by a noncanonical Wnt signaling mediator, duboraya.
2006,
Pubmed Pan,
RhoA-mediated apical actin enrichment is required for ciliogenesis and promoted by Foxj1.
2007,
Pubmed Panizzi,
New functions for a vertebrate Rho guanine nucleotide exchange factor in ciliated epithelia.
2007,
Pubmed Park,
Subcellular localization and signaling properties of dishevelled in developing vertebrate embryos.
2005,
Pubmed
,
Xenbase Park,
Ciliogenesis defects in embryos lacking inturned or fuzzy function are associated with failure of planar cell polarity and Hedgehog signaling.
2006,
Pubmed
,
Xenbase Raji,
Light and electron microscopic studies of the trachea in the one-humped camel (Camelus dromedarius).
2007,
Pubmed Ross,
Disruption of Bardet-Biedl syndrome ciliary proteins perturbs planar cell polarity in vertebrates.
2005,
Pubmed Rothbächer,
Dishevelled phosphorylation, subcellular localization and multimerization regulate its role in early embryogenesis.
2000,
Pubmed
,
Xenbase Scholey,
Intraflagellar transport and cilium-based signaling.
2006,
Pubmed Schwarz-Romond,
The Wnt signalling effector Dishevelled forms dynamic protein assemblies rather than stable associations with cytoplasmic vesicles.
2005,
Pubmed Simons,
Inversin, the gene product mutated in nephronophthisis type II, functions as a molecular switch between Wnt signaling pathways.
2005,
Pubmed
,
Xenbase Sorokin,
Reconstructions of centriole formation and ciliogenesis in mammalian lungs.
1968,
Pubmed Steinman,
An electron microscopic study of ciliogenesis in developing epidermis and trachea in the embryo of Xenopus laevis.
1968,
Pubmed
,
Xenbase Toskala,
Temporal and spatial distribution of ciliogenesis in the tracheobronchial airways of mice.
2005,
Pubmed Vladar,
Molecular characterization of centriole assembly in ciliated epithelial cells.
2007,
Pubmed Wallingford,
Dishevelled controls cell polarity during Xenopus gastrulation.
2000,
Pubmed
,
Xenbase Wallingford,
The developmental biology of Dishevelled: an enigmatic protein governing cell fate and cell polarity.
2005,
Pubmed Wallingford,
Planar cell polarity, ciliogenesis and neural tube defects.
2006,
Pubmed Yu,
Association of Dishevelled with the clathrin AP-2 adaptor is required for Frizzled endocytosis and planar cell polarity signaling.
2007,
Pubmed
,
Xenbase
