In vivo Wnt signaling tracing through a transgenic biosensor fish reveals novel activity domains

Moro, Enrico; Özhan, Günes; Mongera, Alessandro; Beis, Dimitris; Wierzbicki, Claudia; Young, Rodrigo; Bournele, Despina; Domenichini, Alice; Valdivia, Leonardo E; Lum, Lawrence; Chen, Chuo; Amatruda, James F.; Tiso, Natascia; Weidinger, Gilbert; Argenton, Francesco

doi:10.1016/j.ydbio.2012.03.023

Cited by 237 publications

(288 citation statements)

References 67 publications

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“…The following lines were used: AB, TL, Tg(kdrl:GFP) s843 (Jin et al, 2005), Tg(kdrl:ras-mCherry) s896 (Chi et al, 2008), Tg(7xTCF-Xla. Siam:GFP) ia4 (Moro et al, 2012), Tg(-17.0neurog1:EGFP) w61 (McGraw et al, 2008), ouchless (sorbs3 w35 ) (Malmquist et al, 2013), adgra2 s984 and adgra2 s985 (Vanhollebeke et al, 2015), as reported previously. All animal experiments were performed in accordance with the rules of the State of Belgium ( protocol approval number: CEBEA-IBMM-2012:65).…”

Section: Zebrafish Strains and Cell Linessupporting

confidence: 62%

Defective adgra2 (gpr124) splicing and function in zebrafish ouchless mutants

et al. 2016

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A hitherto unidentified N-ethyl-N-nitrosourea (ENU)-induced mutation affects dorsal root ganglia (DRG) formation in ouchless mutant zebrafish larvae. In contrast to previous findings assigning the ouchless phenotypes to downregulated sorbs3 transcript levels, this work re-attributes the phenotypes to an essential splice site mutation affecting adgra2 (gpr124) splicing and function. Accordingly, ouchless mutants fail to complement previously characterized adgra2 mutants and exhibit highly penetrant cerebrovascular defects. The aberrantly spliced adgra2 transcript found in ouchless mutants encodes a receptor lacking a single leucine-rich repeat (LRR) within its N-terminus.

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Section: Zebrafish Strains and Cell Linessupporting

confidence: 62%

Defective adgra2 (gpr124) splicing and function in zebrafish ouchless mutants

et al. 2016

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“…Hence, Wnt3EGFP transgenics might not be ideal for assessing the efficacy of Wnt inhibition by IWR-1. In this respect, these transgenics complement those developed to analyze downstream effects of Wnt signaling (Moro et al, 2012). Nevertheless, the results of IWR-1 treatment emphasize the specificity of C59 action and support the existence of a secreted Wnt3 fraction.…”

Section: Discussionmentioning

confidence: 76%

Modulating expression level of secreted Wnt3 influences cerebellum development in zebrafish transgenics

et al. 2015

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The boundaries of brain regions are associated with the tissuespecific secretion of ligands from different signaling pathways. The dynamics of these ligands in vivo and the impact of its disruption remain largely unknown. Using light and fluorescence microscopy for the overall imaging of the specimen and fluorescence correlation spectroscopy (FCS) to determine Wnt3 dynamics, we demonstrated that Wnt3 regulates cerebellum development during embryogenesis using zebrafish wnt3 transgenics with either tissue-specific expression of an EGFP reporter or a functionally active fusion protein, Wnt3EGFP. The results suggest a state of dynamic equilibrium of Wnt3EGFP mobility in polarized neuroepithelial-like progenitors in the dorsal midline and cerebellar progenitors on the lateral side. Wnt3EGFP is secreted from the cerebellum as shown by measurements of its mobility in the ventricular cavity. The importance of Wnt secretion in brain patterning was validated with the Porcn inhibitor Wnt-C59 (C59), which, when applied early, reduced membrane-bound and secreted fractions of Wnt3EGFP and led to a malformed brain characterized by the absence of epithalamus, optic tectum and cerebellum. Likewise, interference with Wnt secretion later on during cerebellar development negatively impacted cerebellar growth and patterning. Our work, supported by quantitative analysis of protein dynamics in vivo, highlights the importance of membrane-localized and secreted Wnt3 during cerebellum development.

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“…Loss of zygotic β-catenin-dependent Wnt signaling also results in axial defects (Lekven et al, 2001;Agathon et al, 2003;Shimizu et al, 2005;Petersen and Reddien, 2009 (Dorsky et al, 2002;Moro et al, 2012). Although smp knockdown did not affect Tg(7xTCF:mCherry) reporter expression at 85% epiboly (Fig.…”

Section: Introductionmentioning

confidence: 88%

“…Transgenic Wnt reporter line Tg(7xTCF-XLa.Siam:nlsmCherry) ia5 (Kwan et al, 2007;Moro et al, 2012). Other transgenic lines were Tg(Top:dGFP) (Dorsky et al, 2002), Tg(hsp70:dkk1-GFP) and Tg(hsp70:wnt8a-GFP) (Stoick-Cooper et al, 2007), and Tg(hsp70:GFP) (Halloran et al, 2000).…”

Section: Transgenic Linesmentioning

confidence: 99%

Simplet/Fam53b is required for Wnt signal transduction by regulating β-catenin nuclear localization

et al. 2014

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Canonical β-catenin-dependent Wnt signal transduction is important for several biological phenomena, such as cell fate determination, cell proliferation, stem cell maintenance and anterior-posterior axis formation. The hallmark of canonical Wnt signaling is the translocation of β-catenin into the nucleus where it activates gene transcription. However, the mechanisms regulating β-catenin nuclear localization are poorly understood. We show that Simplet/ Fam53B (Smp) is required for Wnt signaling by positively regulating β-catenin nuclear localization. In the zebrafish embryo, the loss of smp blocks the activity of two β-catenin-dependent reporters and the expression of Wnt target genes, and prevents nuclear accumulation of β-catenin. Conversely, overexpression of smp increases β-catenin nuclear localization and transcriptional activity in vitro and in vivo. Expression of mutant Smp proteins lacking either the nuclear localization signal or the β-catenin interaction domain reveal that the translocation of Smp into the nucleus is essential for β-catenin nuclear localization and Wnt signaling in vivo. We also provide evidence that mammalian Smp is involved in regulating β-catenin nuclear localization: the protein colocalizes with β-catenin-dependent gene expression in mouse intestinal crypts; siRNA knockdown of Smp reduces β-catenin nuclear localization and transcriptional activity; human SMP mediates β-catenin transcriptional activity in a dose-dependent manner; and the human SMP protein interacts with human β-catenin primarily in the nucleus. Thus, our findings identify the evolutionary conserved SMP protein as a regulator of β-catenindependent Wnt signal transduction.

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In vivo Wnt signaling tracing through a transgenic biosensor fish reveals novel activity domains

Cited by 237 publications

References 67 publications

Defective adgra2 (gpr124) splicing and function in zebrafish ouchless mutants

Defective adgra2 (gpr124) splicing and function in zebrafish ouchless mutants

Modulating expression level of secreted Wnt3 influences cerebellum development in zebrafish transgenics

Simplet/Fam53b is required for Wnt signal transduction by regulating β-catenin nuclear localization

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