Molecular Physiology, Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.
Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chikusa, Nagoya 464-8601, Japan.
Plant Physiol. 2021 Oct 5;187(2):485-503. doi: 10.1093/plphys/kiab353.
The understanding of signaling and metabolic processes in multicellular organisms requires knowledge of the spatial dynamics of small molecules and the activities of enzymes, transporters, and other proteins in vivo, as well as biophysical parameters inside cells and across tissues. The cellular distribution of receptors, ligands, and activation state must be integrated with information about the cellular distribution of metabolites in relation to metabolic fluxes and signaling dynamics in order to achieve the promise of in vivo biochemistry. Genetically encoded sensors are engineered fluorescent proteins that have been developed for a wide range of small molecules, such as ions and metabolites, or to report biophysical processes, such as transmembrane voltage or tension. First steps have been taken to monitor the activity of transporters in vivo. Advancements in imaging technologies and specimen handling and stimulation have enabled researchers in plant sciences to implement sensor technologies in intact plants. Here, we provide a brief history of the development of genetically encoded sensors and an overview of the types of sensors available for quantifying and visualizing ion and metabolite distribution and dynamics. We further discuss the pros and cons of specific sensor designs, imaging systems, and sample manipulations, provide advice on the choice of technology, and give an outlook into future developments.
多细胞生物中信号和代谢过程的理解需要了解小分子的空间动态以及体内酶、转运蛋白和其他蛋白质的活性，以及细胞内和组织间的生物物理参数。受体、配体和激活状态的细胞分布必须与代谢通量和信号动力学相关的代谢物细胞分布信息相结合，以实现体内生物化学的承诺。遗传编码传感器是为广泛的小分子（如离子和代谢物）或报告生物物理过程（如跨膜电压或张力）而设计的工程化荧光蛋白。已经采取了初步措施来监测体内转运蛋白的活性。成像技术以及标本处理和刺激的进步使植物科学研究人员能够在完整的植物中实施传感器技术。在这里，我们简要介绍了遗传编码传感器的发展历史，并概述了可用于量化和可视化离子和代谢物分布和动态的传感器类型。我们进一步讨论了特定传感器设计、成像系统和样本处理的优缺点，提供了有关技术选择的建议，并展望了未来的发展。
