Pryor SE , Massa V , Savery D , Andre P , Yang Y , Greene ND , Copp AJ .

083361 Wellcome Trust , 087259 Wellcome Trust , 087525 Wellcome Trust , G0801124 Medical Research Council , Wellcome Trust

	Fig. 1. Normal pattern of NC cell migration in Vangl2Lp/Lp mouse embryos. Migrating NC detected by Erbb3 mRNA expression (A-F) and YFP expression regulated by Wnt1-Cre (G-N). Wild-type (+/+; A-C,G-J) and Vangl2Lp/Lp (Lp/Lp; D-F,K-N) embryos at early E9.5 (13-14 somites) both exhibit NC cells colonising forebrain, peri-ocular region (A,D, arrows) and upper branchial arches (ba). Transverse sections show branchial arch colonisation (B,E,H,I,L,M) and migration from closed (+/+) and open (Lp/Lp) neural tube (arrows in C,F,J,N). da, dorsal aorta. Scale bars: 500†µm in A,D; 200†µm in B,C,E,F; 250†µm in G,K; 100†µm in H-J,L-N.
	Fig. 2. Vangl1 is not expressed in wild-type NC, nor ectopically in Vangl2Lp/Lp mutants. WISH in intact embryos shown from dorsal (A,E,I,M) and right lateral (A′,E′,I′,M′) views, and in sections (B-D,F-H,J-L,N-P) at levels indicated by dashed lines in A,E,I,M. Vangl1 mRNA expression is confined to midline neuroepithelium, from hindbrain to low trunk (arrows), in E8.5 wild-type embryos (+/+; A-D). There is no ectopic expression in Vangl2Lp/Lp embryos (Lp/Lp; E-H) nor overlap with Erbb3-positive NC (I-L). Vangl2 is expressed throughout the neuroepithelium (M-P), overlapping with Vangl1 only in midline cells, and not overlapping with Erbb3. Scale bars: 200†µm.
	Fig. 3. Normal NC migration in Vangl1/2 double mutants and after acute Vangl2 downregulation in the NC lineage. (A-H) Control (A,C-E; Vangl1gt/+; Vangl2Δ/+) and double-mutant (B,F-H; Vangl1gt/gt; Vangl2Δ/Δ) embryos exhibit normal migration of Erbb3-positive cranial NC (E8.5; A,B) and cranial/trunk NC (E9.5; C-H). Acute NC downregulation of Vangl2 to test for a possible compensatory mechanism in Vangl2Lp/Lp embryos (I) reveals identical YFP-positive NC migration in control (J-L; Vangl2+/flox; Wnt1-Cre) and downregulation (M-O; Vangl2Lp/flox; Wnt1-Cre) E9.5 embryos. Arrows indicate comparable streams of NC cells migrating from the trunk neural tube in both genotypes. Scale bars: 200†µm in A; 500†µm in C,F; 100†µm in J-O.
	Fig. 4. NC cells migrate similarly from Vangl2+/+ and Vangl2Lp/Lp explant cultures. YFP-positive NC are initially (0†h; A) on the dorsal margin of Vangl2+/+; Wnt1-Cre/YFP (+/+) explants and on Vangl2Lp/Lp; Wnt1-Cre/YFP (Lp/Lp) neural fold tips. Cells emerge in similar numbers (at 24†h; A), with no difference in outgrowth area (P=0.91, one-way ANOVA; B). Leading edge NC cells (anti-GFP/YFP; DAPI) are polarised (C, arrows) or non-polarised (C, arrowheads). Analysis of leading edge cells (D) reveals no difference between genotypes in the proportion of polarised cells nor in the mean distance migrated (P=0.42, E; P=0.21, F; one-way ANOVA). bf, bright field. Error bars indicate s.e.m. At least three explants were studied per genotype and time point. Scale bars: 200†µm in A; 50†µm in C.

Benish, Letter: "The neurocristopathies: a unifying concept of disease arising in neural crest development". 1975, Pubmed

Benish, Letter: "The neurocristopathies: a unifying concept of disease arising in neural crest development". 1975, Pubmed
Carmona-Fontaine, Contact inhibition of locomotion in vivo controls neural crest directional migration. 2008, Pubmed , Xenbase
Cheung, Neural crest development is regulated by the transcription factor Sox9. 2003, Pubmed
De Calisto, Essential role of non-canonical Wnt signalling in neural crest migration. 2005, Pubmed , Xenbase
Dolk, The prevalence of congenital anomalies in Europe. 2010, Pubmed
Doudney, Analysis of the planar cell polarity gene Vangl2 and its co-expressed paralogue Vangl1 in neural tube defect patients. 2005, Pubmed
Etheridge, Murine dishevelled 3 functions in redundant pathways with dishevelled 1 and 2 in normal cardiac outflow tract, cochlea, and neural tube development. 2008, Pubmed
Formstone, The flamingo-related mouse Celsr family (Celsr1-3) genes exhibit distinct patterns of expression during embryonic development. 2001, Pubmed
Garratt, Neuregulin, a factor with many functions in the life of a schwann cell. 2000, Pubmed
Gray, Diversification of the expression patterns and developmental functions of the dishevelled gene family during chordate evolution. 2009, Pubmed , Xenbase
Guirao, Coupling between hydrodynamic forces and planar cell polarity orients mammalian motile cilia. 2010, Pubmed
Hamblet, Dishevelled 2 is essential for cardiac outflow tract development, somite segmentation and neural tube closure. 2002, Pubmed
Harris, Mouse mutants with neural tube closure defects and their role in understanding human neural tube defects. 2007, Pubmed
Henderson, Cardiovascular defects associated with abnormalities in midline development in the Loop-tail mouse mutant. 2001, Pubmed
Ikeya, Wnt signalling required for expansion of neural crest and CNS progenitors. 1997, Pubmed
Jiang, Fate of the mammalian cardiac neural crest. 2000, Pubmed
Juriloff, A consideration of the evidence that genetic defects in planar cell polarity contribute to the etiology of human neural tube defects. 2012, Pubmed
Kibar, Ltap, a mammalian homolog of Drosophila Strabismus/Van Gogh, is altered in the mouse neural tube mutant Loop-tail. 2001, Pubmed
Kulesa, Cranial neural crest migration: new rules for an old road. 2010, Pubmed
Matthews, Directional migration of neural crest cells in vivo is regulated by Syndecan-4/Rac1 and non-canonical Wnt signaling/RhoA. 2008, Pubmed , Xenbase
Merte, Sec24b selectively sorts Vangl2 to regulate planar cell polarity during neural tube closure. 2010, Pubmed
Montcouquiol, Identification of Vangl2 and Scrb1 as planar polarity genes in mammals. 2003, Pubmed
Murdoch, Severe neural tube defects in the loop-tail mouse result from mutation of Lpp1, a novel gene involved in floor plate specification. 2001, Pubmed
Sauka-Spengler, A gene regulatory network orchestrates neural crest formation. 2008, Pubmed
Seifert, Frizzled/PCP signalling: a conserved mechanism regulating cell polarity and directed motility. 2007, Pubmed
Shima, Differential expression of the seven-pass transmembrane cadherin genes Celsr1-3 and distribution of the Celsr2 protein during mouse development. 2002, Pubmed
Shnitsar, PTK7 recruits dsh to regulate neural crest migration. 2008, Pubmed , Xenbase
Song, Planar cell polarity breaks bilateral symmetry by controlling ciliary positioning. 2010, Pubmed
Srinivas, Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. 2001, Pubmed
Torban, Tissue, cellular and sub-cellular localization of the Vangl2 protein during embryonic development: effect of the Lp mutation. 2007, Pubmed
Torban, Genetic interaction between members of the Vangl family causes neural tube defects in mice. 2008, Pubmed
Torban, Independent mutations in mouse Vangl2 that cause neural tube defects in looptail mice impair interaction with members of the Dishevelled family. 2004, Pubmed
Vandenberg, Non-canonical Wnt signaling regulates cell polarity in female reproductive tract development via van gogh-like 2. 2009, Pubmed
Yates, The PCP genes Celsr1 and Vangl2 are required for normal lung branching morphogenesis. 2010, Pubmed
Yates, The planar cell polarity gene Vangl2 is required for mammalian kidney-branching morphogenesis and glomerular maturation. 2010, Pubmed
Ybot-Gonzalez, Convergent extension, planar-cell-polarity signalling and initiation of mouse neural tube closure. 2007, Pubmed
Yin, Comparison of phenotypes between different vangl2 mutants demonstrates dominant effects of the Looptail mutation during hair cell development. 2012, Pubmed