Luo Jian, Wyss-Coray Tony
Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA, USA.
Methods Mol Biol. 2009;574:193-202. doi: 10.1007/978-1-60327-321-3_16.
TGF-beta signaling via the Smad2/3 pathway has key roles in development and tissue homeostasis. Perturbations of the TGF-beta 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.
通过Smad2/3途径的转化生长因子β（TGF-β）信号传导在发育和组织稳态中起关键作用。TGF-β信号传导的扰动涉及许多人类疾病的发病机制，包括癌症、纤维化疾病、发育缺陷和神经退行性变。为了研究活体动物中Smad2/3依赖性信号传导的时空模式，我们构建了带有Smad反应性荧光素酶报告基因的转基因小鼠（SBE-luc小鼠）。通过生物发光成像可以在活体小鼠中无创地评估Smad2/3依赖性信号传导。为了确定生物发光信号的细胞来源，我们构建了新的报告基因小鼠，其在相同的SBE启动子控制下表达一种包含荧光素酶、红色荧光蛋白（RFP）和胸苷激酶的三融合蛋白（SBE-lucRT小鼠）。SBE-luc和SBE-lucRT小鼠可用于研究活体小鼠中Smad2/3依赖性信号传导的时间、组织特异性激活，以及用于鉴定该途径的内源性或合成调节剂。
