Genetics of Development and Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD, 20892, USA.
Vascular Biology Program, Boston Children's Hospital, Department of Surgery, Harvard Medical School, Boston, MA, 02115, USA.
Nat Commun. 2017 Oct 27;8(1):1163. doi: 10.1038/s41467-017-01340-7.
G protein-coupled receptors (GPCRs), a superfamily of cell-surface receptors involved in virtually all physiological processes, are the major target class for approved drugs. Imaging GPCR activation in real time in living animals would provide a powerful way to study their role in biology and disease. Here, we describe a mouse model that enables the bioluminescent detection of GPCR activation in real time by utilizing the clinically important GPCR, sphingosine-1-phosphate receptor 1 (S1P). A synthetic S1P signaling pathway, designed to report the interaction between S1P and β-arrestin2 via the firefly split luciferase fragment complementation system, is genetically encoded in these mice. Upon receptor activation and subsequent β-arrestin2 recruitment, an active luciferase enzyme complex is produced, which can be detected by in vivo bioluminescence imaging. This imaging strategy reveals the dynamics and spatial specificity of S1P activation in normal and pathophysiologic contexts in vivo and can be applied to other GPCRs.
G 蛋白偶联受体（GPCRs）是细胞表面受体的超家族，参与几乎所有的生理过程，是已批准药物的主要靶标类。在活体动物中实时成像 GPCR 的激活将为研究它们在生物学和疾病中的作用提供一种强大的方法。在这里，我们描述了一种小鼠模型，该模型通过利用临床重要的 GPCR 鞘氨醇-1-磷酸受体 1（S1P），可以实时进行生物发光检测 GPCR 的激活。设计用于通过萤火虫分裂荧光素酶片段互补系统报告 S1P 与β-arrestin2 相互作用的合成 S1P 信号通路，在这些小鼠中进行基因编码。在受体激活和随后的β-arrestin2 募集之后，产生活性荧光素酶酶复合物，可通过体内生物发光成像进行检测。这种成像策略揭示了 S1P 在正常和病理生理情况下在体内的激活的动态和空间特异性，并且可以应用于其他 GPCR。
