Whitaker Michael
Institute of Cell and Molecular Biosciences Medical School, Newcastle University, Framlington Place Newcastle upon Tyne, United Kingdom.
Methods Cell Biol. 2010;99:153-82. doi: 10.1016/B978-0-12-374841-6.00006-2.
Small, fluorescent, calcium-sensing molecules have been enormously useful in mapping intracellular calcium signals in time and space, as chapters in this volume attest. Despite their widespread adoption and utility, they suffer some disadvantages. Genetically encoded calcium sensors that can be expressed inside cells by transfection or transgenesis are desirable. The last 10 years have been marked by a rapid evolution in the laboratory of genetically encoded calcium sensors both figuratively and literally, resulting in 11 distinct configurations of fluorescent proteins and their attendant calcium sensor modules. Here, the design logic and performance of this abundant collection of sensors and their in vitro and in vivo use and performance are described. Genetically encoded calcium sensors have proved valuable in the measurement of calcium concentration in cellular organelles, for the most part in single cells in vitro. Their success as quantitative calcium sensors in tissues in vitro and in vivo is qualified, but they have proved valuable in imaging the pattern of calcium signals within tissues in whole animals. Some branches of the calcium sensor evolutionary tree continue to evolve rapidly and the steady progress in optimizing sensor parameters leads to the certain hope that these drawbacks will eventually be overcome by further genetic engineering.
正如本书各章节所证明的那样，小型荧光钙传感分子在绘制细胞内钙信号的时空分布方面非常有用。尽管它们被广泛采用且具有实用性，但也存在一些缺点。通过转染或转基因在细胞内表达的基因编码钙传感器是很有必要的。在过去的十年里，基因编码钙传感器在实验室里经历了迅速的发展，无论是从字面意义还是比喻意义上来说都是如此，产生了11种不同构型的荧光蛋白及其相关的钙传感器模块。在此，将描述这大量传感器的设计逻辑、性能以及它们在体外和体内的使用与性能。基因编码钙传感器已被证明在测量细胞器中的钙浓度方面很有价值，在很大程度上是用于体外的单细胞。它们作为体外和体内组织中定量钙传感器的成功是有限的，但已被证明在对整个动物组织内钙信号模式进行成像方面很有价值。钙传感器进化树的一些分支仍在迅速进化，并且在优化传感器参数方面的稳步进展带来了一定的希望，即这些缺点最终将通过进一步的基因工程得以克服。
