Generation and application of signaling pathway reporter lines in zebrafish

Moro, Enrico; Vettori, Andrea; Porazzi, Patrizia; Schiavone, Marco; Rampazzo, Elena; Casari, Alessandro; Ek, Olivier; Facchinello, Nicola; Astone, Matteo; Zancan, Ilaria; Milanetto, Martina; Tiso, Natascia; Argenton, Francesco

doi:10.1007/s00438-013-0750-z

Cited by 68 publications

(59 citation statements)

References 62 publications

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“…Endothelial: Tg(kdrl:RFP) s896 (Chi et al, 2008), Tg(kdrl:EGFP) la116 (Anderson et al, 2008) Tg(kdrl:eGFP-NLS) zf109 (Blum et al, 2008), Tg(fli1a:eGFP) y1 ; erythrocytes: Tg(gata1a:DsRed) sd2 (Traver et al, 2003); DRG (and other neuronal populations): Tg(neurog1: eGFP) w61 (McGraw et al, 2008); canonical Wnt signaling activity: Tg(7xTCF-Xla.Siam:GFP) ia4 (Moro et al, 2013). Transgenes for inhibiting canonical Wnt signaling were Tg(hsp70l:Mmu.Axin1-YFP) w35 (Kagermeier-Schenk et al, 2011) and Tg(hsp70l:Xla.TCFΔC-EGFP) (Martin and Kimelman, 2012).…”

Section: Transgenesmentioning

confidence: 99%

Reck enables cerebrovascular development by promoting canonical Wnt signaling

et al. 2015

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The cerebral vasculature provides the massive blood supply that the brain needs to grow and survive. By acquiring distinctive cellular and molecular characteristics it becomes the blood-brain barrier (BBB), a selectively permeable and protective interface between the brain and the peripheral circulation that maintains the extracellular milieu permissive for neuronal activity. Accordingly, there is great interest in uncovering the mechanisms that modulate the formation and differentiation of the brain vasculature. By performing a forward genetic screen in zebrafish we isolated no food for thought (nft y72 ), a recessive late-lethal mutant that lacks most of the intracerebral central arteries (CtAs), but not other brain blood vessels. We found that the cerebral vascularization deficit of nft y72 mutants is caused by an inactivating lesion in reversion-inducing cysteine-rich protein with Kazal motifs [reck; also known as suppressor of tumorigenicity 15 protein (ST15)], which encodes a membrane-anchored tumor suppressor glycoprotein. Our findings highlight Reck as a novel and pivotal modulator of the canonical Wnt signaling pathway that acts in endothelial cells to enable intracerebral vascularization and proper expression of molecular markers associated with BBB formation. Additional studies with cultured endothelial cells suggest that, in other contexts, Reck impacts vascular biology via the vascular endothelial growth factor (VEGF) cascade. Together, our findings have broad implications for both vascular and cancer biology.

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Section: Transgenesmentioning

confidence: 99%

Reck enables cerebrovascular development by promoting canonical Wnt signaling

et al. 2015

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“…S4) (Moro et al, 2013). According to what has already been shown ex vivo on human pancreatic cancer (Bailey and Leach, 2012; De La O et al, 2008; Hu et al, 2012), we confirmed in vivo the upregulation of the Notch signaling pathway during tumor onset (Fig.…”

Section: Discussionmentioning

confidence: 99%

Zebrafish reporter lines reveal in vivo signaling pathway activities involved in pancreatic cancer

Schiavone

Rampazzo

Casari

et al. 2014

Disease Models &Amp; Mechanisms

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Pancreatic adenocarcinoma, one of the worst malignancies of the exocrine pancreas, is a solid tumor with increasing incidence and mortality in industrialized countries. This condition is usually driven by oncogenic KRAS point mutations and evolves into a highly aggressive metastatic carcinoma due to secondary gene mutations and unbalanced expression of genes involved in the specific signaling pathways. To examine in vivo the effects of KRASG12D during pancreatic cancer progression and time correlation with cancer signaling pathway activities, we have generated a zebrafish model of pancreatic adenocarcinoma in which eGFP-KRASG12D expression was specifically driven to the pancreatic tissue by using the GAL4/UAS conditional expression system. Outcrossing the inducible oncogenic KRASG12D line with transgenic zebrafish reporters, harboring specific signaling responsive elements of transcriptional effectors, we were able to follow TGFβ, Notch, Bmp and Shh activities during tumor development. Zebrafish transgenic lines expressing eGFP-KRASG12D showed normal exocrine pancreas development until 3 weeks post fertilization (wpf). From 4 to 24 wpf we observed several degrees of acinar lesions, characterized by an increase in mesenchymal cells and mixed acinar/ductal features, followed by progressive bowel and liver infiltrations and, finally, highly aggressive carcinoma. Moreover, live imaging analysis of the exocrine pancreatic tissue revealed an increasing number of KRAS-positive cells and progressive activation of TGFβ and Notch pathways. Increase in TGFβ, following KRASG12D activation, was confirmed in a concomitant model of medulloblastoma (MDB). Notch and Shh signaling activities during tumor onset were different between MDB and pancreatic adenocarcinoma, indicating a tissue-specific regulation of cell signaling pathways. Moreover, our results show that a living model of pancreatic adenocarcinoma joined with cell signaling reporters is a suitable tool for describing in vivo the signaling cascades and molecular mechanisms involved in tumor development and a potential platform to screen for novel oncostatic drugs.

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“…In addition to tools for the visualization of cellular morphological changes, an increasing panel of signaling pathway reporter lines is now available for live monitoring of intracellular signaling activities [reviewed in [46]]. These biosensor lines have been successfully used to monitor the activity of BMP, FGF, sonic hedgehog, Wnt and Notch pathways in zebrafish embryos [47,48,49,50,51].…”

Section: The Transgenic Toolkit For Zebrafish Thyroid Organogenesismentioning

confidence: 99%

New Model Systems to Illuminate Thyroid Organogenesis. Part I: An Update on the Zebrafish Toolbox

2013

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Thyroid dysgenesis (TD) resulting from defects during embryonic thyroid development represents a major cause of congenital hypothyroidism. The pathogenetic mechanisms of TD in human newborns, however, are still poorly understood and disease-causing genetic variants have been identified in only a small percentage of TD cases. This limited understanding of the pathogenesis of TD is partly due to a lack of knowledge on how intrinsic factors and extrinsic signalling cues orchestrate the differentiation of thyroid follicular cells and the morphogenesis of thyroid tissue. Recently, embryonic stem cells and zebrafish embryos emerged as novel model systems that allow for innovative experimental approaches in order to decipher cellular and molecular mechanisms of thyroid development and to unravel pathogenic mechanisms of TD. Zebrafish embryos offer several salient properties for studies on thyroid organogenesis including rapid and external development, optical transparency, ease of breeding, relative short generation time and amenability for genome editing. In this review, we will highlight recent advances in the zebrafish toolkit to visualize cellular dynamics of organ development and discuss specific prospects of the zebrafish model for studies on vertebrate thyroid development and human congenital thyroid diseases.

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Generation and application of signaling pathway reporter lines in zebrafish

Cited by 68 publications

References 62 publications

Reck enables cerebrovascular development by promoting canonical Wnt signaling

Reck enables cerebrovascular development by promoting canonical Wnt signaling

Zebrafish reporter lines reveal in vivo signaling pathway activities involved in pancreatic cancer

New Model Systems to Illuminate Thyroid Organogenesis. Part I: An Update on the Zebrafish Toolbox

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