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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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Organization of the apical MT cytoskeleton in early third-instar wing discs.(A) Tubulin staining in third-instar wing pouch. Note the asymmetric accumulation of tubulin on the proximal side of the cell in early third-instar wing pouch. Scale bar, 2.5 Δm. White arrows show the proximal distal axis in the wing disc, with distal in the center of the wing pouch, and proximal tissue forming a ring around the future wing blade. (A′) Correlation analysis of image (A) for tubulin and Dsh. Note the increasing overlap of signal when the image of tubulin is shifted leftward (proximal) relative to the Dsh image, resulting in a peak at distance A. A rightward shift (distal) initially shows a decrease in correlation of tubulin and Dsh signals, with a peak at larger distance B, indicating that, on average, the tubulin signals are closer to the proximal than to the distal sides of cells. Similar dorsal or ventral translocations show that tubulin signals are, on average, equidistant between dorsal and ventral sides of the cell. (B–D) Orientations of apical MTs correlate with core PCP protein polarization and Fj/Ds gradients throughout wing development. Spatiotemporal correlation of core PCP protein polarity (Dsh::GFP) with MT alignment (TyrTUB) at time points from 19 to 30 hAPF. Rose plots show distributions of MT orientation (P-D axis, corresponding to the Fj and Ds gradients, is plotted as horizontal (90°); Plots, derived from OrientationJ, are composed of 36 bins of 5° each). Note that orientation remains polarized until 30 hAPF, at which time orientations become randomized. Scale bars: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.005
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fig1s1: Organization of the apical MT cytoskeleton in early third-instar wing discs.(A) Tubulin staining in third-instar wing pouch. Note the asymmetric accumulation of tubulin on the proximal side of the cell in early third-instar wing pouch. Scale bar, 2.5 Δm. White arrows show the proximal distal axis in the wing disc, with distal in the center of the wing pouch, and proximal tissue forming a ring around the future wing blade. (A′) Correlation analysis of image (A) for tubulin and Dsh. Note the increasing overlap of signal when the image of tubulin is shifted leftward (proximal) relative to the Dsh image, resulting in a peak at distance A. A rightward shift (distal) initially shows a decrease in correlation of tubulin and Dsh signals, with a peak at larger distance B, indicating that, on average, the tubulin signals are closer to the proximal than to the distal sides of cells. Similar dorsal or ventral translocations show that tubulin signals are, on average, equidistant between dorsal and ventral sides of the cell. (B–D) Orientations of apical MTs correlate with core PCP protein polarization and Fj/Ds gradients throughout wing development. Spatiotemporal correlation of core PCP protein polarity (Dsh::GFP) with MT alignment (TyrTUB) at time points from 19 to 30 hAPF. Rose plots show distributions of MT orientation (P-D axis, corresponding to the Fj and Ds gradients, is plotted as horizontal (90°); Plots, derived from OrientationJ, are composed of 36 bins of 5° each). Note that orientation remains polarized until 30 hAPF, at which time orientations become randomized. Scale bars: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.005

Mentions: Apical MT alignment and orientation of core PCP protein domains have been shown to correlate, but have only been examined in several small domains during late pupal wing development (Shimada et al., 2006; Harumoto et al., 2010) (between 14 and 30 hr after puparium formation [APF]) (See also Figure 1—figure supplement 1B–D). If MT alignment provides directional bias for core protein polarization, one should observe a spatiotemporal correlation across the entire wing throughout the time core PCP proteins are polarized.


Microtubules provide directional information for core PCP function.

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

Organization of the apical MT cytoskeleton in early third-instar wing discs.(A) Tubulin staining in third-instar wing pouch. Note the asymmetric accumulation of tubulin on the proximal side of the cell in early third-instar wing pouch. Scale bar, 2.5 Δm. White arrows show the proximal distal axis in the wing disc, with distal in the center of the wing pouch, and proximal tissue forming a ring around the future wing blade. (A′) Correlation analysis of image (A) for tubulin and Dsh. Note the increasing overlap of signal when the image of tubulin is shifted leftward (proximal) relative to the Dsh image, resulting in a peak at distance A. A rightward shift (distal) initially shows a decrease in correlation of tubulin and Dsh signals, with a peak at larger distance B, indicating that, on average, the tubulin signals are closer to the proximal than to the distal sides of cells. Similar dorsal or ventral translocations show that tubulin signals are, on average, equidistant between dorsal and ventral sides of the cell. (B–D) Orientations of apical MTs correlate with core PCP protein polarization and Fj/Ds gradients throughout wing development. Spatiotemporal correlation of core PCP protein polarity (Dsh::GFP) with MT alignment (TyrTUB) at time points from 19 to 30 hAPF. Rose plots show distributions of MT orientation (P-D axis, corresponding to the Fj and Ds gradients, is plotted as horizontal (90°); Plots, derived from OrientationJ, are composed of 36 bins of 5° each). Note that orientation remains polarized until 30 hAPF, at which time orientations become randomized. Scale bars: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.005
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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fig1s1: Organization of the apical MT cytoskeleton in early third-instar wing discs.(A) Tubulin staining in third-instar wing pouch. Note the asymmetric accumulation of tubulin on the proximal side of the cell in early third-instar wing pouch. Scale bar, 2.5 Δm. White arrows show the proximal distal axis in the wing disc, with distal in the center of the wing pouch, and proximal tissue forming a ring around the future wing blade. (A′) Correlation analysis of image (A) for tubulin and Dsh. Note the increasing overlap of signal when the image of tubulin is shifted leftward (proximal) relative to the Dsh image, resulting in a peak at distance A. A rightward shift (distal) initially shows a decrease in correlation of tubulin and Dsh signals, with a peak at larger distance B, indicating that, on average, the tubulin signals are closer to the proximal than to the distal sides of cells. Similar dorsal or ventral translocations show that tubulin signals are, on average, equidistant between dorsal and ventral sides of the cell. (B–D) Orientations of apical MTs correlate with core PCP protein polarization and Fj/Ds gradients throughout wing development. Spatiotemporal correlation of core PCP protein polarity (Dsh::GFP) with MT alignment (TyrTUB) at time points from 19 to 30 hAPF. Rose plots show distributions of MT orientation (P-D axis, corresponding to the Fj and Ds gradients, is plotted as horizontal (90°); Plots, derived from OrientationJ, are composed of 36 bins of 5° each). Note that orientation remains polarized until 30 hAPF, at which time orientations become randomized. Scale bars: 5 μm.DOI:http://dx.doi.org/10.7554/eLife.02893.005
Mentions: Apical MT alignment and orientation of core PCP protein domains have been shown to correlate, but have only been examined in several small domains during late pupal wing development (Shimada et al., 2006; Harumoto et al., 2010) (between 14 and 30 hr after puparium formation [APF]) (See also Figure 1—figure supplement 1B–D). If MT alignment provides directional bias for core protein polarization, one should observe a spatiotemporal correlation across the entire wing throughout the time core PCP proteins are polarized.

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