European Molecular Biology Laboratory (EMBL), Barcelona, Spain.
Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.
Methods Mol Biol. 2022;2525:321-332. doi: 10.1007/978-1-0716-2473-9_25.
Somites are formed sequentially by the segmentation of the anterior parts of the presomitic mesoderm (PSM), and such periodical somite formation is crucial to ensure the proper vertebrae. In the mouse embryo, Hes7, a segmentation clock gene, controls this periodic event with new somites forming every 2 h. Hes7 oscillations are synchronized between neighboring PSM cells and propagate from the posterior to the anterior PSM in the form of traveling waves. However, the exact mechanisms that generate these oscillatory dynamics and control synchronization are still unclear. Given that the half-life of Hes7 is too short to be monitored with most fluorescent proteins, time-lapse bioluminescence imaging (BLI) is a suitable tool to monitor the chronological Hes7 expression dynamics. In this chapter, we introduce a ubiquitinated luciferase reporter which enables the visualization of Hes7 expression dynamics with high temporal and spatial resolution in living cells and tissues.
体节是由前节间中胚层（PSM）的分段形成的，这种周期性的体节形成对于确保适当的脊椎骨至关重要。在小鼠胚胎中，分割时钟基因 Hes7 控制着这个周期性事件，每隔 2 小时就会形成新的体节。Hes7 振荡在相邻 PSM 细胞之间同步，并以传播波的形式从前部向后部 PSM 传播。然而，产生这些振荡动力学并控制同步的确切机制仍不清楚。鉴于 Hes7 的半衰期太短，无法用大多数荧光蛋白进行监测，因此延时生物发光成像（BLI）是监测 Hes7 表达动态的合适工具。在本章中，我们介绍了一种泛素化的荧光素酶报告基因，它能够以高时空分辨率在活细胞和组织中可视化 Hes7 的表达动态。
