title: Syndecan 4 interacts genetically with Vangl2 to regulate neural tube closure and planar cell polarity.
abstract: Syndecan 4 (Sdc4) is a cell-surface heparan sulfate proteoglycan (HSPG) that regulates gastrulation, neural tube closure and directed neural crest migration in Xenopus development. To determine whether Sdc4 participates in Wnt/PCP signaling during mouse development, we evaluated a possible interaction between a null mutation of Sdc4 and the loop-tail allele of Vangl2. Sdc4 is expressed in multiple tissues, but particularly in the non-neural ectoderm, hindgut and otic vesicles. Sdc4;Vangl2(Lp) compound mutant mice have defective spinal neural tube closure, disrupted orientation of the stereocilia bundles in the cochlea and delayed wound healing, demonstrating a strong genetic interaction. In Xenopus, co-injection of suboptimal amounts of Sdc4 and Vangl2 morpholinos resulted in a significantly greater proportion of embryos with defective neural tube closure than each individual morpholino alone. To probe the mechanism of this interaction, we overexpressed or knocked down Vangl2 function in HEK293 cells. The Sdc4 and Vangl2 proteins colocalize, and Vangl2, particularly the Vangl2(Lp) mutant form, diminishes Sdc4 protein levels. Conversely, Vangl2 knockdown enhances Sdc4 protein levels. Overall HSPG steady-state levels were regulated by Vangl2, suggesting a molecular mechanism for the genetic interaction in which Vangl2(Lp/+) enhances the Sdc4-null phenotype. This could be mediated via heparan sulfate residues, as Vangl2(Lp/+) embryos fail to initiate neural tube closure and develop craniorachischisis (usually seen only in Vangl2(Lp/Lp)) when cultured in the presence of chlorate, a sulfation inhibitor. These results demonstrate that Sdc4 can participate in the Wnt/PCP pathway, unveiling its importance during neural tube closure in mammalian embryos.
authors.affiliation: Center for Aging and Regeneration, Millennium Nucleus in Regenerative Biology, Faculty of Biological Sciences, P. Universidad Católica de Chile, Alameda 340 Santiago, Chile.
authors.forename: Noelia
authors.initials: N
authors.lastname: Escobedo
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Contreras
authors.forename: Rosana
authors.initials: R
authors.lastname: Muñoz
authors.forename: Marjorie
authors.initials: M
authors.lastname: Farías
authors.forename: Héctor
authors.initials: H
authors.lastname: Carrasco
authors.forename: Charlotte
authors.initials: C
authors.lastname: Hill
authors.forename: Uyen
authors.initials: U
authors.lastname: Tran
authors.forename: Sophie E
authors.initials: SE
authors.lastname: Pryor
authors.forename: Oliver
authors.initials: O
authors.lastname: Wessely
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Copp
authors.forename: Juan
authors.initials: J
authors.lastname: Larraín
mesh_terms: D000818:Animals; D016764:Cell Polarity; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005260:Female; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006198:Hair Cells, Auditory / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D054259:Neural Tube / Q000166:cytology* / Q000378:metabolism; D009436:Neural Tube Defects / Q000378:metabolism; D053671:Syndecan-4 / Q000235:genetics / Q000378:metabolism*; D060449:Wnt Signaling Pathway; D014945:Wound Healing; D014981:Xenopus
chemical_list: D006497:Heparan Sulfate; D009419:Nerve Tissue Proteins; D053671:Syndecan-4; C431577:Ltap protein, mouse; C508306:Sdc4 protein, mouse
keywords: Neural tube defects; Proteoglycans; Wnt planar cell polarity