PUBLICATION
A Smad3 transgenic reporter reveals TGF-beta control of zebrafish spinal cord development
- Authors
- Casari, A., Schiavone, M., Facchinello, N., Vettori, A., Meyer, D., Tiso, N., Moro, E., Argenton, F.
- ID
- ZDB-PUB-141007-1
- Date
- 2014
- Source
- Developmental Biology 396(1): 81-93 (Journal)
- Registered Authors
- Argenton, Francesco, Casari, Alessandro, Facchinello, Nicola, Meyer, Dirk, Moro, Enrico, Schiavone, Marco, Tiso, Natascia, Vettori, Andrea
- Keywords
- Proliferation, Reporter, Smad3, TGFbeta, Transcription, Transgenic, Zebrafish
- MeSH Terms
-
- Gene Expression Regulation, Developmental*
- Transforming Growth Factor beta/metabolism*
- Zebrafish
- Cell Proliferation
- Spinal Cord/embryology*
- Spinal Cord/physiology
- Transgenes*
- Cell Cycle
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- Protein Serine-Threonine Kinases/metabolism
- Signal Transduction
- Phenotype
- Animals
- Immunohistochemistry
- Activin Receptors, Type I/metabolism
- Receptors, Transforming Growth Factor beta/metabolism
- Neurons/metabolism
- Genes, Reporter
- Promoter Regions, Genetic
- Animals, Genetically Modified
- Smad3 Protein/genetics*
- PubMed
- 25286120 Full text @ Dev. Biol.
Citation
Casari, A., Schiavone, M., Facchinello, N., Vettori, A., Meyer, D., Tiso, N., Moro, E., Argenton, F. (2014) A Smad3 transgenic reporter reveals TGF-beta control of zebrafish spinal cord development. Developmental Biology. 396(1):81-93.
Abstract
TGF-beta (TGFβ) family mediated Smad signaling is involved in mesoderm and endoderm specification, left-right asymmetry formation and neural tube development. The TGFβ1/2/3 and Activin/Nodal signal transduction cascades culminate with activation of SMAD2 and/or SMAD3 transcription factors and their overactivation are involved in different pathologies with an inflammatory and/or uncontrolled cell proliferation basis, such as cancer and fibrosis. We have developed a transgenic zebrafish reporter line responsive to Smad3 activity. Through chemical, genetic and molecular approaches we have seen that this transgenic line consistently reproduces in vivo Smad3-mediated TGFβ signaling. Reporter fluorescence is activated in phospho-Smad3 positive cells and is responsive to both Smad3 isoforms, Smad3a and 3b. Moreover, Alk4 and Alk5 inhibitors strongly repress the reporter activity. In the CNS, Smad3 reporter activity is particularly high in the subpallium, tegumentum, cerebellar plate, medulla oblongata and the retina proliferative zone. In the spinal cord, the reporter is activated at the ventricular zone, where neuronal progenitor cells are located. Colocalization methods show in vivo that TGFβ signaling is particularly active in neuroD+ precursors. Using neuronal transgenic lines, we observed that TGFβ chemical inhibition leads to a decrease of differentiating cells and an increase of proliferation. Similarly, smad3a and 3b knock-down alter neural differentiation showing that both paralogues play a positive role in neural differentiation. EdU proliferation assay and pH3 staining confirmed that Smad3 is mainly active in post-mitotic, non-proliferating cells. In summary, we demonstrate that the Smad3 reporter line allows us to follow in vivo Smad3 transcriptional activity and that Smad3, by controlling neural differentiation, promotes the progenitor to precursor switch allowing neural progenitors to exit cell cycle and differentiate.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping