scholarly journals A new transgenic reporter line reveals Wnt-dependent Snail2 reexpression and cranial neural crest differentiation in Xenopus

2019 ◽  
Author(s):  
Jiejing Li ◽  
Mark Perfetto ◽  
Christopher Materna ◽  
Rebecca Li ◽  
Hong Thi Tran ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Fluorescent Protein ◽  
Developmental Stages ◽  
Egfp Expression ◽  
Cranial Neural Crest ◽  
Canonical Wnt Signaling ◽  
Reporter Line ◽  
Green Fluorescent ◽  
Canonical Wnt

AbstractDuring vertebrate embryogenesis, the cranial neural crest (CNC) forms at the neural plate border and subsequently migrates and differentiates into many types of cells. The transcription factor Snail2, which is induced by canonical Wnt signaling to be expressed in the early CNC, is pivotal for CNC induction and migration in Xenopus. However, snail2 expression is silenced during CNC migration, and its roles at later developmental stages remain unclear. We generated a transgenic X. tropicalis line that expresses enhanced green fluorescent protein (eGFP) driven by the snail2 promoter/enhancer, and observed eGFP expression not only in the pre-migratory and migrating CNC, but also the differentiating CNC. This transgenic line can be used directly to detect deficiencies in CNC development at various stages, including subtle perturbation of CNC differentiation. In situ hybridization and immunohistochemistry confirm that Snail2 is reexpressed in the differentiating CNC. Using a separate transgenic Wnt reporter line, we show that canonical Wnt signaling is also active in the differentiating CNC. Blocking Wnt signaling shortly after CNC migration causes reduced snail2 expression and impaired differentiation of CNC-derived head cartilage structures. These results suggest that Wnt signaling drives the reexpression of snail2 in the post-migratory CNC and regulates CNC differentiation.

scholarly journals Draxin acts as a molecular rheostat of canonical Wnt signaling to control cranial neural crest EMT

2018 ◽  
Vol 217 (10) ◽  
pp. 3683-3697 ◽  
Author(s):  
Erica J. Hutchins ◽  
Marianne E. Bronner
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Neural Tube ◽  
Epithelial To Mesenchymal Transition ◽  
Cranial Neural Crest ◽  
Canonical Wnt Signaling ◽  
Mesenchymal Transition ◽  
Molecular Rheostat ◽  
Timing Of Initiation ◽  
Canonical Wnt

Neural crest cells undergo a spatiotemporally regulated epithelial-to-mesenchymal transition (EMT) that proceeds head to tailward to exit from the neural tube. In this study, we show that the secreted molecule Draxin is expressed in a transient rostrocaudal wave that mirrors this emigration pattern, initiating after neural crest specification and being down-regulated just before delamination. Functional experiments reveal that Draxin regulates the timing of cranial neural crest EMT by transiently inhibiting canonical Wnt signaling. Ectopic maintenance of Draxin in the cranial neural tube blocks full EMT; while cells delaminate, they fail to become mesenchymal and migratory. Loss of Draxin results in premature delamination but also in failure to mesenchymalize. These results suggest that a pulse of intermediate Wnt signaling triggers EMT and is necessary for its completion. Taken together, these data show that transient secreted Draxin mediates proper levels of canonical Wnt signaling required to regulate the precise timing of initiation and completion of cranial neural crest EMT.

scholarly journals Transcriptomic Identification of Draxin-Responsive Targets During Cranial Neural Crest EMT

2021 ◽  
Vol 12 ◽  
Author(s):  
Erica J. Hutchins ◽  
Michael L. Piacentino ◽  
Marianne E. Bronner
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Gene List ◽  
Hybridization Chain Reaction ◽  
Cranial Neural Crest ◽  
Canonical Wnt Signaling ◽  
Chain Reaction ◽  
Mesenchymal Transition ◽  
Neural Crest Development ◽  
Canonical Wnt

Canonical Wnt signaling plays an essential role in proper craniofacial morphogenesis, at least partially due to regulation of various aspects of cranial neural crest development. In an effort to gain insight into the etiology of craniofacial abnormalities resulting from Wnt signaling and/or cranial neural crest dysfunction, we sought to identify Wnt-responsive targets during chick cranial neural crest development. To this end, we leveraged overexpression of a canonical Wnt antagonist, Draxin, in conjunction with RNA-sequencing of cranial neural crest cells that have just activated their epithelial–mesenchymal transition (EMT) program. Through differential expression analysis, gene list functional annotation, hybridization chain reaction (HCR), and quantitative reverse transcription polymerase chain reaction (RT-qPCR), we validated a novel downstream target of canonical Wnt signaling in cranial neural crest – RHOB – and identified possible signaling pathway crosstalk underlying cranial neural crest migration. The results reveal novel putative targets of canonical Wnt signaling during cranial neural crest EMT and highlight important intersections across signaling pathways involved in craniofacial development.

scholarly journals Genetic interaction of Pax3 mutation and canonical Wnt signaling modulates neural tube defects and neural crest abnormalities

genesis ◽  
2021 ◽  
Author(s):  
Alexandra J. Palmer ◽  
Dawn Savery ◽  
Valentina Massa ◽  
Andrew J. Copp ◽  
Nicholas D. E. Greene
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Neural Tube ◽  
Neural Tube Defects ◽  
Genetic Interaction ◽  
Canonical Wnt Signaling ◽  
Canonical Wnt

scholarly journals Enhanced Activation of Canonical Wnt Signaling Confers Mesoderm-Derived Parietal Bone with Similar Osteogenic and Skeletal Healing Capacity to Neural Crest-Derived Frontal Bone

PLoS ONE ◽  
2015 ◽  
Vol 10 (10) ◽  
pp. e0138059 ◽  
Author(s):  
Shuli Li ◽  
Natalina Quarto ◽  
Kshemendra Senarath-Yapa ◽  
Nathaniel Grey ◽  
Xue Bai ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Parietal Bone ◽  
Frontal Bone ◽  
Canonical Wnt Signaling ◽  
Canonical Wnt

scholarly journals Caudal-related Homeobox (Cdx) Protein-dependent Integration of Canonical Wnt Signaling on Paired-box 3 (Pax3) Neural Crest Enhancer

2012 ◽  
Vol 287 (20) ◽  
pp. 16623-16635 ◽  
Author(s):  
Oraly Sanchez- Ferras ◽  
Baptiste Coutaud ◽  
Taraneh Djavanbakht Samani ◽  
Isabelle Tremblay ◽  
Ouliana Souchkova ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Neural Crest ◽  
Canonical Wnt Signaling ◽  
Canonical Wnt

scholarly journals Fluorescence-based gene reporter plasmid to track canonical Wnt signaling in ENS inflammation

2016 ◽  
Vol 310 (6) ◽  
pp. G337-G346
Author(s):  
Rosa Di Liddo ◽  
Thomas Bertalot ◽  
Anne Schuster ◽  
Sandra Schrenk ◽  
Oliver Müller ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Fluorescent Protein ◽  
Reporter Plasmid ◽  
Experimental Conditions ◽  
Highly Sensitive ◽  
Vitro Model ◽  
Green Fluorescent ◽  
Canonical Wnt ◽  
Compelling Experimental Evidence

In several gut inflammatory or cancer diseases, cell-cell interactions are compromised, and an increased cytoplasmic expression of β-catenin is observed. Over the last decade, numerous studies provided compelling experimental evidence that the loss of cadherin-mediated cell adhesion can promote β-catenin release and signaling without any specific activation of the canonical Wnt pathway. In the present work, we took advantage of the ability of lipofectamine-like reagent to cause a synchronous dissociation of adherent junctions in cells isolated from the rat enteric nervous system (ENS) for obtaining an in vitro model of deregulated β-catenin signaling. Under these experimental conditions, a green fluorescent protein Wnt reporter plasmid called ΔTop_EGFP3a was successfully tested to screen β-catenin stabilization at resting and primed conditions with exogenous Wnt3a or lipopolysaccharide (LPS). ΔTop_EGFP3a provided a reliable and strong fluorescent signal that was easily measurable and at the same time highly sensitive to modulations of Wnt signaling following Wnt3a and LPS stimulation. The reporter gene was useful to demonstrate that Wnt3a exerts a protective activity in the ENS from overstimulated Wnt signaling by promoting a downregulation of the total β-catenin level. Based on this evidence, the use of ΔTop_EGFP3a reporter plasmid could represent a more reliable tool for the investigation of Wnt and cross-talking pathways in ENS inflammation.

A novel role for MuSK and non-canonical Wnt signaling during segmental neural crest cell migration

2011 ◽  
Vol 124 (15) ◽  
pp. e1-e1 ◽  
Author(s):  
S. Banerjee ◽  
L. Gordon ◽  
T. M. Donn ◽  
C. Berti ◽  
C. B. Moens ◽  
...  
Keyword(s):  
Cell Migration ◽  
Wnt Signaling ◽  
Neural Crest ◽  
Neural Crest Cell ◽  
Canonical Wnt Signaling ◽  
Neural Crest Cell Migration ◽  
Canonical Wnt ◽  
Crest Cell
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