title: In vivo imaging of Hedgehog pathway activation with a nuclear fluorescent reporter.
abstract: 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.
authors: Department of Biochemistry, Stanford University School of Medicine, Stanford, California, United States of America. Current address: Children's Research Institute, University of Texas-Southwestern Medical Center, Dallas, Texas, United States of America.
authors: John K
authors: JK
authors: Mich
authors: Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors: Alexander Y
authors: AY
authors: Payumo
authors: Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors: Paul G
authors: PG
authors: Rack
authors: Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California, United States of America; Department of Developmental Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors: James K
authors: JK
authors: Chen
mesh_terms: D058500:Animal Fins / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D002467:Cell Nucleus / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D053823:Hedgehog Proteins / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009154:Mutation; D015203:Reproducibility of Results; D015398:Signal Transduction; D019170:Somites / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D059008:Time-Lapse Imaging / Q000379:methods; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000254:growth & development; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D053823:Hedgehog Proteins; D008164:Luminescent Proteins; D029961:Zebrafish Proteins; D000097573:Red Fluorescent Protein; C524053:Shha protein, zebrafish