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Microtubules provide directional information for core PCP function.

Matis M, Russler-Germain DA, Hu Q, Tomlin CJ, Axelrod JD - Elife (2014)

Bottom Line: Consistent with previous results, we find that the Ft/Ds/Fj-module has an effect on a MT-cytoskeleton.We show Ft/Ds/Fj-dependent initial polarization of the apical MT-cytoskeleton prior to global alignment of the core-module, reveal that the anchoring of apical non-centrosomal MTs at apical junctions is polarized, observe that directional trafficking of vesicles containing Dsh depends on Ft, and demonstrate the feasibility of this model by mathematical simulation.Together, these results support the hypothesis that Ft/Ds/Fj provides a signal to orient core PCP function via MT polarization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, United States matism@uni-muenster.de.

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Correlation between EB1 comets and MT orientation in Drosophila wing epithelium.(A and B) Overlays of 84 frames from time-lapse videos (Video 1) in 24 hAPF pupal wing expressing EB1::GFP and Jupiter::Cherry (Jupiter is a MT associated protein). (A′ and B′) Orientations of MTs and EB1 comets (panels on the left) are color-coded using OrientationJ (18). Scale bar: 5 μm. (C) MT staining with anti-α-tubulin antibody is showing identical pattern as staining with anti-Tyr-tubulin antibody. Scale bar: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.006
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fig1s2: Correlation between EB1 comets and MT orientation in Drosophila wing epithelium.(A and B) Overlays of 84 frames from time-lapse videos (Video 1) in 24 hAPF pupal wing expressing EB1::GFP and Jupiter::Cherry (Jupiter is a MT associated protein). (A′ and B′) Orientations of MTs and EB1 comets (panels on the left) are color-coded using OrientationJ (18). Scale bar: 5 μm. (C) MT staining with anti-α-tubulin antibody is showing identical pattern as staining with anti-Tyr-tubulin antibody. Scale bar: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.006

Mentions: Core PCP protein polarization with respect to the tissue axes is first observed during larval wing development (Classen et al., 2005). We therefore surveyed apical MT structure beginning in third instar. To facilitate this broad analysis, we used tubulin staining. While foregoing the ability to determine plus-end orientation as provided by analysis of EB1 comets, this approach enables analysis of vastly greater numbers of MTs than does the EB1 assay, and also provides the potential to distinguish a more stable, anchored population, though in fact we see a strong correlation between results from both methods (Figure 1—figure supplement 2A–B′; Video 1). Similarly, both anti-tubulin and anti-tyrosinated tubulin antibodies produce indistinguishable results (Figure 1—figure supplement 2C).Video 1.Live in vivo imaging of EB1::GFP and Cherry::Jupiter.


Microtubules provide directional information for core PCP function.

Matis M, Russler-Germain DA, Hu Q, Tomlin CJ, Axelrod JD - Elife (2014)

Correlation between EB1 comets and MT orientation in Drosophila wing epithelium.(A and B) Overlays of 84 frames from time-lapse videos (Video 1) in 24 hAPF pupal wing expressing EB1::GFP and Jupiter::Cherry (Jupiter is a MT associated protein). (A′ and B′) Orientations of MTs and EB1 comets (panels on the left) are color-coded using OrientationJ (18). Scale bar: 5 μm. (C) MT staining with anti-α-tubulin antibody is showing identical pattern as staining with anti-Tyr-tubulin antibody. Scale bar: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.006
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4151085&req=5

fig1s2: Correlation between EB1 comets and MT orientation in Drosophila wing epithelium.(A and B) Overlays of 84 frames from time-lapse videos (Video 1) in 24 hAPF pupal wing expressing EB1::GFP and Jupiter::Cherry (Jupiter is a MT associated protein). (A′ and B′) Orientations of MTs and EB1 comets (panels on the left) are color-coded using OrientationJ (18). Scale bar: 5 μm. (C) MT staining with anti-α-tubulin antibody is showing identical pattern as staining with anti-Tyr-tubulin antibody. Scale bar: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.006
Mentions: Core PCP protein polarization with respect to the tissue axes is first observed during larval wing development (Classen et al., 2005). We therefore surveyed apical MT structure beginning in third instar. To facilitate this broad analysis, we used tubulin staining. While foregoing the ability to determine plus-end orientation as provided by analysis of EB1 comets, this approach enables analysis of vastly greater numbers of MTs than does the EB1 assay, and also provides the potential to distinguish a more stable, anchored population, though in fact we see a strong correlation between results from both methods (Figure 1—figure supplement 2A–B′; Video 1). Similarly, both anti-tubulin and anti-tyrosinated tubulin antibodies produce indistinguishable results (Figure 1—figure supplement 2C).Video 1.Live in vivo imaging of EB1::GFP and Cherry::Jupiter.

Bottom Line: Consistent with previous results, we find that the Ft/Ds/Fj-module has an effect on a MT-cytoskeleton.We show Ft/Ds/Fj-dependent initial polarization of the apical MT-cytoskeleton prior to global alignment of the core-module, reveal that the anchoring of apical non-centrosomal MTs at apical junctions is polarized, observe that directional trafficking of vesicles containing Dsh depends on Ft, and demonstrate the feasibility of this model by mathematical simulation.Together, these results support the hypothesis that Ft/Ds/Fj provides a signal to orient core PCP function via MT polarization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, United States matism@uni-muenster.de.

Show MeSH
Related in: MedlinePlus