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

Authors

  • Meng-Chieh Shen,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
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    • Drs. Shen and Ozacar contributed equally to this work.

  • A. Tuba Ozacar,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
    Current affiliation:
    1. Department of Molecular Biology and Genetics, Bogazici University, Istanbul/Turkiye.
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    • Drs. Shen and Ozacar contributed equally to this work.

  • Marcey Osgood,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
    Current affiliation:
    1. Department of Neurology, University of Massachusetts Medical School, Worcester, MA
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  • Crina Boeras,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
    Current affiliation:
    1. Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester, MA
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  • Jochebed Pink,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
    Current affiliation:
    1. Department of Internal Medicine, Emory University Medical School, Atlanta, GA
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  • Jeanne Thomas,

    1. Department of Biology, University of Massachusetts, Amherst, Massachusetts
    Current affiliation:
    1. Department of Genetics, Sunrise Medical Laboratories, Inc., Hicksville, NY
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  • Jhumku D. Kohtz,

    1. Department of Pediatrics, Feinberg School of Medicine, Northwestern University & Lurie Children's Research Center, Chicago, Illinois
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  • Rolf Karlstrom

    Corresponding author
    • Department of Biology, University of Massachusetts, Amherst, Massachusetts
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Correspondence to: R.O. Karlstrom, Department of Biology, University of Massachusetts, Amherst, MA 01003. E-mail: karlstrom@bio.umass.edu

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. Developmental Dynamics 242:529–539, 2013. © 2013 Wiley Periodicals, Inc.

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