Hahn K, DeBiasio R, Taylor D L
Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Nature. 1992 Oct 22;359(6397):736-8. doi: 10.1038/359736a0.
The temporal and spatial dynamics of intracellular signals and protein effectors are being defined as a result of imaging using fluorescent reagents within living cells. We have described a new class of fluorescent analogues termed optical biosensors, which sense chemical or molecular events through their effects on protein transducers. One example of this new class of indicators is MeroCaM, an environmentally sensitive fluorophore which when it is attached to calmodulin reflects the activation of calmodulin by calcium in vitro. We report here that the rise in free calcium and MeroCaM activation occur in the same period during serum stimulation of quiescent fibroblasts. MeroCaM activation also correlates with the spatial pattern of increased free calcium and the contraction of transverse fibres during wound healing. Finally, migrating fibroblasts in the later stages of wound-healing exhibit an increasing gradient of free calcium and MeroCaM activation from the front to the rear.
通过在活细胞内使用荧光试剂进行成像，细胞内信号和蛋白质效应器的时空动态正在被明确。我们描述了一类新的荧光类似物，称为光学生物传感器，它们通过对蛋白质传感器的作用来感知化学或分子事件。这类新指示剂的一个例子是MeroCaM，一种对环境敏感的荧光团，当它与钙调蛋白结合时，在体外反映钙对钙调蛋白的激活。我们在此报告，在血清刺激静止成纤维细胞期间，游离钙的升高和MeroCaM的激活发生在同一时期。MeroCaM的激活还与伤口愈合过程中游离钙增加的空间模式以及横向纤维的收缩相关。最后，在伤口愈合后期迁移的成纤维细胞表现出从前端到后端游离钙和MeroCaM激活的梯度增加。
