Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Dev Dyn. 2011 Mar;240(3):712-22. doi: 10.1002/dvdy.22567. Epub 2011 Feb 8.
Bone morphogenic protein (BMP) signaling is fundamental to development, injury response, and homeostasis. We have developed transgenic zebrafish that report Smad-mediated BMP signaling in embryos and adults. These lines express either enhanced green fluorescent protein (eGFP), destabilized eGFP, or destabilized Kusabira Orange 2 (KO2) under the well-characterized BMP Response Element (BRE). These fluorescent proteins were found to be expressed dynamically in regions of known BMP signaling including the developing tail bud, hematopoietic lineage, dorsal eye, brain structures, heart, jaw, fins, and somites, as well as other tissues. Responsiveness to changes in BMP signaling was confirmed by observing fluorescence after activation in an hsp70:bmp2b transgenic background or by inhibition in an hsp70:nog3 background. We further demonstrated faithful reportage by the BRE transgenic lines following chemical repression of BMP signaling using an inhibitor of BMP receptor activity, dorsomorphin. Overall, these lines will serve as valuable tools to explore the mechanisms and regulation of BMP signal during embryogenesis, in tissue maintenance, and during disease.
骨形态发生蛋白 (BMP) 信号转导对于胚胎发育、损伤反应和内稳态至关重要。我们开发了一种转基因斑马鱼,该鱼能够报告胚胎和成鱼中 Smad 介导的 BMP 信号转导。这些品系在特征明确的 BMP 反应元件 (BRE) 下表达增强型绿色荧光蛋白 (eGFP)、不稳定 eGFP 或不稳定 Kusabira Orange 2 (KO2)。这些荧光蛋白在已知的 BMP 信号转导区域(包括正在发育的尾芽、造血谱系、背眼、脑结构、心脏、下颚、鳍和体节以及其他组织)中动态表达。在 hsp70:bmp2b 转基因背景下观察到激活后的荧光或在 hsp70:nog3 背景下观察到抑制后的荧光,证实了对 BMP 信号转导变化的反应性。我们进一步通过使用 BMP 受体活性抑制剂 dorsomorphin 抑制 BMP 信号转导,证明了 BRE 转基因系的忠实报告。总体而言,这些系将成为探索胚胎发生、组织维持和疾病期间 BMP 信号转导机制和调控的有价值的工具。