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Sulf1 has ligand-dependent effects on canonical and non-canonical Wnt signalling.

Fellgett SW, Maguire RJ, Pownall ME - J. Cell. Sci. (2015)

Bottom Line: Sulf1 could, therefore, influence the formation of Wnt signalling complexes to modulate the activation of both canonical and non-canonical pathways.In addition, we model the ability of Sulf1 to influence morphogen gradients using fluorescently tagged Wnt ligands in ectodermal explants.We show that Sulf1 overexpression has ligand-specific effects on Wnt signalling: it affects membrane accumulation and extracellular levels of tagged Wnt8a and Wnt11b ligands differently, and inhibits the activity of canonical Wnt8a but enhances the activity of non-canonical Wnt11b.

View Article: PubMed Central - PubMed

Affiliation: Biology Department, University of York, York YO10 5YW, UK.

No MeSH data available.


Sulf1 enhances the secretion and range of diffusion of Wnt8a–GFP in animal explants. (A) Diagram depicting the assay used to measure Wnt8a–GFP secretion and diffusion through a control background, see Materials and Methods for details. (B,C) mRNA encoding either (B) mCerulean (600 pg), LacZ (4 ng) and Wnt8a–GFP (2 ng) or (C) mRFP (600 pg), Sulf1 (4 ng) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. (D) The range of diffusion of Wnt8a–GFP through a control background was quantified using Fiji Image J. (E) Diagram depicting the assay used to measure Wnt8a–GFP diffusion through a background expressing Sulf1. (F–I) mRNA encoding mCerulean (600 pg) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. An adjacent blastomere was injected with mRNA encoding (F,G) mRFP (600 pg) and LacZ (4 ng) or (H,I) mRFP (600 pg) and Sulf1 (4 ng). (J) The range of Wnt8a–GFP through a background expressing either LacZ or Sulf1 was quantified using Fiji Image J. Scale bars: 20 μm.
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f07: Sulf1 enhances the secretion and range of diffusion of Wnt8a–GFP in animal explants. (A) Diagram depicting the assay used to measure Wnt8a–GFP secretion and diffusion through a control background, see Materials and Methods for details. (B,C) mRNA encoding either (B) mCerulean (600 pg), LacZ (4 ng) and Wnt8a–GFP (2 ng) or (C) mRFP (600 pg), Sulf1 (4 ng) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. (D) The range of diffusion of Wnt8a–GFP through a control background was quantified using Fiji Image J. (E) Diagram depicting the assay used to measure Wnt8a–GFP diffusion through a background expressing Sulf1. (F–I) mRNA encoding mCerulean (600 pg) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. An adjacent blastomere was injected with mRNA encoding (F,G) mRFP (600 pg) and LacZ (4 ng) or (H,I) mRFP (600 pg) and Sulf1 (4 ng). (J) The range of Wnt8a–GFP through a background expressing either LacZ or Sulf1 was quantified using Fiji Image J. Scale bars: 20 μm.

Mentions: We tested the ability of Sulf1 to influence Wnt8a diffusion when co-expressed in the ligand-producing cell. At the four-cell stage, a single animal blastomere was microinjected with mRNAs encoding Wnt8a–GFP together with a membrane marker and either Sulf1 or LacZ (as a control) (Fig. 7A). The level of GFP fluorescence was measured across the field of cells, with mCerulean marking the source of the fluorescent Wnt ligand. We found that in control conditions, Wnt8a–GFP was capable of diffusing two or three cell diameters away from the cells expressing it (Fig. 7B). Co-expression of Sulf1 significantly increased the range of Wnt8a–GFP diffusion to ∼six or seven cell diameters (Fig. 7C). The effects of Sulf1 on Wnt8a–GFP diffusion for multiple experiments were quantified (Fig. 7D). The graph shows the reduction in the average fluorescence intensity of Wnt8a–GFP with increasing distance from Wnt secreting cells. Sulf1 increased the levels of Wnt8a–GFP further from the source as compared to controls (Fig. 7D). The lines of best fit for the Wnt8a–GFP distribution show similar rates of decay under both control and Sulf1 conditions. One prediction from this is that Sulf1 does not alter the qualitative nature of Wnt8a–GFP diffusion, but rather enhances the levels of Wnt8a–GFP released from Wnt-secreting cells, allowing increased range of signalling.


Sulf1 has ligand-dependent effects on canonical and non-canonical Wnt signalling.

Fellgett SW, Maguire RJ, Pownall ME - J. Cell. Sci. (2015)

Sulf1 enhances the secretion and range of diffusion of Wnt8a–GFP in animal explants. (A) Diagram depicting the assay used to measure Wnt8a–GFP secretion and diffusion through a control background, see Materials and Methods for details. (B,C) mRNA encoding either (B) mCerulean (600 pg), LacZ (4 ng) and Wnt8a–GFP (2 ng) or (C) mRFP (600 pg), Sulf1 (4 ng) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. (D) The range of diffusion of Wnt8a–GFP through a control background was quantified using Fiji Image J. (E) Diagram depicting the assay used to measure Wnt8a–GFP diffusion through a background expressing Sulf1. (F–I) mRNA encoding mCerulean (600 pg) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. An adjacent blastomere was injected with mRNA encoding (F,G) mRFP (600 pg) and LacZ (4 ng) or (H,I) mRFP (600 pg) and Sulf1 (4 ng). (J) The range of Wnt8a–GFP through a background expressing either LacZ or Sulf1 was quantified using Fiji Image J. Scale bars: 20 μm.
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Related In: Results  -  Collection

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f07: Sulf1 enhances the secretion and range of diffusion of Wnt8a–GFP in animal explants. (A) Diagram depicting the assay used to measure Wnt8a–GFP secretion and diffusion through a control background, see Materials and Methods for details. (B,C) mRNA encoding either (B) mCerulean (600 pg), LacZ (4 ng) and Wnt8a–GFP (2 ng) or (C) mRFP (600 pg), Sulf1 (4 ng) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. (D) The range of diffusion of Wnt8a–GFP through a control background was quantified using Fiji Image J. (E) Diagram depicting the assay used to measure Wnt8a–GFP diffusion through a background expressing Sulf1. (F–I) mRNA encoding mCerulean (600 pg) and Wnt8a–GFP (2 ng) was injected into the animal hemisphere of one blastomere at the four-cell stage. An adjacent blastomere was injected with mRNA encoding (F,G) mRFP (600 pg) and LacZ (4 ng) or (H,I) mRFP (600 pg) and Sulf1 (4 ng). (J) The range of Wnt8a–GFP through a background expressing either LacZ or Sulf1 was quantified using Fiji Image J. Scale bars: 20 μm.
Mentions: We tested the ability of Sulf1 to influence Wnt8a diffusion when co-expressed in the ligand-producing cell. At the four-cell stage, a single animal blastomere was microinjected with mRNAs encoding Wnt8a–GFP together with a membrane marker and either Sulf1 or LacZ (as a control) (Fig. 7A). The level of GFP fluorescence was measured across the field of cells, with mCerulean marking the source of the fluorescent Wnt ligand. We found that in control conditions, Wnt8a–GFP was capable of diffusing two or three cell diameters away from the cells expressing it (Fig. 7B). Co-expression of Sulf1 significantly increased the range of Wnt8a–GFP diffusion to ∼six or seven cell diameters (Fig. 7C). The effects of Sulf1 on Wnt8a–GFP diffusion for multiple experiments were quantified (Fig. 7D). The graph shows the reduction in the average fluorescence intensity of Wnt8a–GFP with increasing distance from Wnt secreting cells. Sulf1 increased the levels of Wnt8a–GFP further from the source as compared to controls (Fig. 7D). The lines of best fit for the Wnt8a–GFP distribution show similar rates of decay under both control and Sulf1 conditions. One prediction from this is that Sulf1 does not alter the qualitative nature of Wnt8a–GFP diffusion, but rather enhances the levels of Wnt8a–GFP released from Wnt-secreting cells, allowing increased range of signalling.

Bottom Line: Sulf1 could, therefore, influence the formation of Wnt signalling complexes to modulate the activation of both canonical and non-canonical pathways.In addition, we model the ability of Sulf1 to influence morphogen gradients using fluorescently tagged Wnt ligands in ectodermal explants.We show that Sulf1 overexpression has ligand-specific effects on Wnt signalling: it affects membrane accumulation and extracellular levels of tagged Wnt8a and Wnt11b ligands differently, and inhibits the activity of canonical Wnt8a but enhances the activity of non-canonical Wnt11b.

View Article: PubMed Central - PubMed

Affiliation: Biology Department, University of York, York YO10 5YW, UK.

No MeSH data available.