Heat-shock-mediated conditional regulation of hedgehog/gli signaling in zebrafish

Dev Dyn. 2013 May;242(5):539-49. doi: 10.1002/dvdy.23955. Epub 2013 Mar 27.

Abstract

Background: Hedgehog (Hh) signaling is required for embryogenesis and continues to play key roles postembryonically in many tissues, influencing growth, stem cell proliferation, and tumorigenesis. Systems for conditional regulation of Hh signaling facilitate the study of these postembryonic 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 hr 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- to 12-hr pulse of Hh signal activation or inactivation, respectively. Using both in situ hybridization and quantitative polymerase chain reaction, 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 central nervous system.

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

Publication types

  • Research Support, N.I.H., Extramural

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 Proteins / genetics
  • Zebrafish Proteins / metabolism
  • Zebrafish* / embryology
  • Zebrafish* / genetics
  • Zebrafish* / metabolism
  • Zinc Finger Protein GLI1

Substances

  • HSP70 Heat-Shock Proteins
  • Hedgehog Proteins
  • Oncogene Proteins
  • Trans-Activators
  • Zebrafish Proteins
  • Zinc Finger Protein GLI1
  • Green Fluorescent Proteins