Protocol

Bioluminescence

Volume 574 of the series Methods in Molecular Biology™ pp 193-202

Date:

Bioluminescence Analysis of Smad-Dependent TGF-β Signaling in Live Mice

  • Jian LuoAffiliated withDepartment of Neurology and Neurological Sciences, Stanford University School of Medicine
  • , Tony Wyss-CorayAffiliated withDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineGRECC, VA Palo Alto Health Care System

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Abstract

TGF-β signaling via the Smad2/3 pathway has key roles in development and tissue homeostasis. Perturbations of the TGF-β signaling are involved in the pathogenesis of many human diseases, including cancer, fibrotic disorders, developmental defects, and neurodegeneration. To study the temporal and spatial patterns of Smad2/3-dependent signaling in living animals, we engineered transgenic mice with a Smad-responsive luciferase reporter (SBE-luc mice). Smad2/3-dependent signaling can be assessed non-invasively in living mice by bioluminescence imaging. To identify the cellular source of the bioluminescence signal, we generated new reporter mice expressing a trifusion protein containing luciferase, red fluorescent protein (RFP), and thymidine kinase under the control of the same SBE promoter (SBE-lucRT mice). SBE-luc and SBE-lucRT mice can be used to study temporal, tissue-specific activation of Smad2/3-dependent signaling in living mice as well as for the identification of endogenous or synthetic modulators of this pathway.

Key words

TGF-β Smad bioluminescence imaging in vivo luciferase immunofluorescence