title: Planar cell polarity proteins differentially regulate extracellular matrix organization and assembly during zebrafish gastrulation.
abstract: Zebrafish gastrulation cell movements occur in the context of dynamic changes in extracellular matrix (ECM) organization and require the concerted action of planar cell polarity (PCP) proteins that regulate cell elongation and mediolateral alignment. Data obtained using Xenopus laevis gastrulae have shown that integrin-fibronectin interactions underlie the formation of polarized cell protrusions necessary for PCP and have implicated PCP proteins themselves as regulators of ECM. By contrast, the relationship between establishment of PCP and ECM assembly/remodeling during zebrafish gastrulation is unclear. We previously showed that zebrafish embryos carrying a null mutation in the four-pass transmembrane PCP protein vang-like 2 (vangl2) exhibit increased matrix metalloproteinase activity and decreased immunolabeling of fibronectin. These data implicated for the first time a core PCP protein in the regulation of pericellular proteolysis of ECM substrates and raised the question of whether other zebrafish PCP proteins also impact ECM organization. In Drosophila melanogaster, the cytoplasmic PCP protein Prickle binds Van Gogh and regulates its function. Here we report that similar to vangl2, loss of zebrafish prickle1a decreases fibronectin protein levels in gastrula embryos. We further show that Prickle1a physically binds Vangl2 and regulates both the subcellular distribution and total protein level of Vangl2. These data suggest that the ability of Prickle1a to impact fibronectin organization is at least partly due to effects on Vangl2. In contrast to loss of either Vangl2 or Prickle1a function, we find that glypican4 (a Wnt co-receptor) and frizzled7 mutant gastrula embryos with disrupted non-canonical Wnt signaling exhibit the opposite phenotype, namely increased fibronectin assembly. Our data show that glypican4 mutants do not have decreased proteolysis of ECM substrates, but instead have increased cell surface cadherin protein expression and increased intercellular adhesion. These data indicate that Wnt/Glypican4/Frizzled signaling regulates ECM assembly through effects on cadherin-mediated cell cohesion. Together, our results demonstrate that zebrafish Vangl2/Prickle1a and non-canonical Wnt/Frizzled signaling have opposing effects on ECM organization underlying PCP and gastrulation cell movements.
authors.affiliation: Division of Genetic Medicine/Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Dohn
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Mundell
authors.forename: Leah M
authors.initials: LM
authors.lastname: Sawyer
authors.forename: Julie A
authors.initials: JA
authors.lastname: Dunlap
authors.forename: Jason R
authors.initials: JR
authors.lastname: Jessen
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D016764:Cell Polarity / Q000502:physiology*; D005109:Extracellular Matrix / Q000502:physiology*; D054262:Gastrulation / Q000502:physiology*; D055785:Gene Knockdown Techniques; D053673:Glypicans / Q000378:metabolism; D047468:Immunoprecipitation; D060588:LIM Domain Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D011956:Receptors, Cell Surface / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000502:physiology*; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D053673:Glypicans; D060588:LIM Domain Proteins; D008565:Membrane Proteins; C558598:Prickle1a protein, zebrafish; D011956:Receptors, Cell Surface; D029961:Zebrafish Proteins; C470289:fzd7a protein, zebrafish; C491759:vangl2 protein, zebrafish
keywords: Cadherins; Fibronectin; Gastrulation; Metalloproteinases; Polarity; Zebrafish