Open AccessIn Vivo Imaging of Hedgehog Pathway Activation with a Nuclear Fluorescent Reporter
Author(s) -
John K. Mich,
Alexander Y. Payumo,
Philip D. Rack,
James Chen
Publication year - 2014
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0103661
Subject(s) - hedgehog , zebrafish , hedgehog signaling pathway , mcherry , microbiology and biotechnology , biology , transcription factor , transgene , live cell imaging , green fluorescent protein , signal transduction , genetics , cell , gene
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.