In vivo imaging of Hedgehog pathway activation with a nuclear fluorescent reporter

The Hedgehog (Hh) pathway is essential for embryonic development and tissue regeneration, and its dysregulation can lead to birth defects and tumorigenesis. Understanding how this signaling mechanism contributes to these processes would benefit from an ability to visualize Hedgehog pathway activity...

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Bibliographic Details
Published in:PloS one 2014-07, Vol.9 (7), p.e103661-e103661
Main Authors: Mich, John K, Payumo, Alexander Y, Rack, Paul G, Chen, James K
Format: Article
Language:English
Subjects:
Animal Fins - embryology
Animal Fins - growth & development
Animal Fins - metabolism
Animal genetic engineering
Animals
Animals, Genetically Modified
Binding sites
Biology
Biology and Life Sciences
Birth defects
Cell Nucleus - metabolism
Congenital defects
Danio rerio
Dihydrofolate reductase
Embryogenesis
Embryonic growth stage
Fluorescence
Genetic research
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Hedgehog protein
Hedgehog Proteins - genetics
Hedgehog Proteins - metabolism
Journalists
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Medicine
Microscopy, Fluorescence
Mutation
Proteins
Red Fluorescent Protein
Regeneration
Regulators
Reproducibility of Results
Research and Analysis Methods
Rodents
Signal Transduction
Signaling
Somites - embryology
Somites - growth & development
Somites - metabolism
Time-Lapse Imaging - methods
Tissue engineering
Transgenic animals
Tumorigenesis
Vertebrates
Zebrafish
Zebrafish - embryology
Zebrafish - genetics
Zebrafish - growth & development
Zebrafish Proteins - genetics
Zebrafish Proteins - metabolism
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Summary:The Hedgehog (Hh) pathway is essential for embryonic development and tissue regeneration, and its dysregulation can lead to birth defects and tumorigenesis. Understanding how this signaling mechanism contributes to these processes would benefit from an ability to visualize Hedgehog pathway activity in live organisms, in real time, and with single-cell resolution. We report here the generation of transgenic zebrafish lines that express nuclear-localized mCherry fluorescent protein in a Gli transcription factor-dependent manner. As demonstrated by chemical and genetic perturbations, these lines faithfully report Hedgehog pathway state in individual cells and with high detection sensitivity. They will be valuable tools for studying dynamic Gli-dependent processes in vertebrates and for identifying new chemical and genetic regulators of the Hh pathway.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0103661