ZFIN ID: ZDB-PUB-130312-13
Heat-shock–mediated conditional regulation of hedgehog/gli signaling in zebrafish
Shen, M., Ozacar, A.T., Osgood, M., Boeras, C., Pink, J., Thomas, J., Kohtz, J., and Karlstrom, R.O.
Date: 2013
Source: Developmental dynamics : an official publication of the American Association of Anatomists   242(5): 539-49 (Journal)
Registered Authors: Karlstrom, Rolf, Osgood, Marcey, Ozacar, Ayse Tuba, Thomas, Jeanne
Keywords: Sonic Hedgehog, Gli, heat shock, transgenic, zebrafish
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Embryo, Nonmammalian
  • Gene Expression Regulation, Developmental/genetics
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • HSP70 Heat-Shock Proteins/genetics
  • HSP70 Heat-Shock Proteins/metabolism
  • Heat-Shock Response/genetics
  • Heat-Shock Response/physiology*
  • Hedgehog Proteins/genetics*
  • Hedgehog Proteins/metabolism
  • Oncogene Proteins/genetics*
  • Oncogene Proteins/metabolism
  • Promoter Regions, Genetic/genetics
  • Signal Transduction/genetics
  • Trans-Activators/genetics*
  • Trans-Activators/metabolism
  • Zebrafish*/embryology
  • Zebrafish*/genetics
  • Zebrafish*/metabolism
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed: 23441066 Full text @ Dev. Dyn.
FIGURES
ABSTRACT

Background: Hedgehog (Hh) signaling is required for embryogenesis and continues to play key roles post-embryonically in many tissues, influencing growth, stem cell proliferation, and tumorigenesis. Systems for conditional regulation of Hh signaling facilitate the study of these post-embryonic Hh functions.

Results: We used the hsp70l promoter to generated three heat-shock-inducible transgenic lines that activate Hh signaling and one line that represses Hh signaling. Heat-shock activation of these transgenes appropriately recapitulates early embryonic loss or gain of Hh function phenotypes. Hh signaling remains activated 24 hours after heat shock in the Tg(hsp70l:shha-EGFP) and Tg(hsp70l:dnPKA-BGFP) lines, while a single heat shock of the Tg(hsp70l:gli1-EGFP) or Tg(hsp70l:gli2aDR-EGFP) lines results in a 6-12 hour pulse of Hh signal activation or inactivation, respectively. Using both in situ hybridization and quantitative PCR, we show that these lines can be used to manipulate Hh signaling through larval and juvenile stages. A ptch2 promoter element was used to generate new reporter lines that allow clear visualization of Hh responding cells throughout the life cycle, including graded Hh responses in the embryonic CNS.

Conclusions: These zebrafish transgenic lines provide important new experimental tools to study the embryonic and post-embryonic roles of Hh signaling.

ADDITIONAL INFORMATION