Rose Tobias, Goltstein Pieter M, Portugues Ruben, Griesbeck Oliver
Max-Planck-Institute of Neurobiology Martinsried, Germany.
Front Mol Neurosci. 2014 Nov 18;7:88. doi: 10.3389/fnmol.2014.00088. eCollection 2014.
More than a decade ago genetically encoded calcium indicators (GECIs) entered the stage as new promising tools to image calcium dynamics and neuronal activity in living tissues and designated cell types in vivo. From a variety of initial designs two have emerged as promising prototypes for further optimization: FRET (Förster Resonance Energy Transfer)-based sensors and single fluorophore sensors of the GCaMP family. Recent efforts in structural analysis, engineering and screening have broken important performance thresholds in the latest generation for both classes. While these improvements have made GECIs a powerful means to perform physiology in living animals, a number of other aspects of sensor function deserve attention. These aspects include indicator linearity, toxicity and slow response kinetics. Furthermore creating high performance sensors with optically more favorable emission in red or infrared wavelengths as well as new stably or conditionally GECI-expressing animal lines are on the wish list. When the remaining issues are solved, imaging of GECIs will finally have crossed the last milestone, evolving from an initial promise into a fully matured technology.
十多年前，基因编码钙指示剂（GECIs）作为一种新的有前景的工具登上舞台，用于对活体组织中的钙动力学和神经元活动以及体内特定细胞类型进行成像。从最初的各种设计中，有两种已成为进一步优化的有前景的原型：基于荧光共振能量转移（FRET）的传感器和GCaMP家族的单荧光团传感器。最近在结构分析、工程和筛选方面的努力突破了这两类传感器最新一代的重要性能阈值。虽然这些改进使GECIs成为在活体动物中进行生理学研究的有力手段，但传感器功能的许多其他方面也值得关注。这些方面包括指示剂线性、毒性和缓慢的反应动力学。此外，创建在红色或红外波长具有更有利光学发射的高性能传感器以及新的稳定或条件性表达GECI的动物品系也在愿望清单上。当剩下的问题得到解决时，GECIs成像最终将跨越最后一个里程碑，从最初的承诺发展成为一项完全成熟的技术。
