Department of Pharmacology, University of California, San Diego, CA, USA.
Shu Chien-Gene Lay Department of Bioengineering, University of California, San Diego, CA, USA.
J Physiol. 2023 Oct;601(19):4227-4241. doi: 10.1113/JP282696. Epub 2023 Sep 25.
Cells execute specific responses to diverse environmental cues by encoding information in distinctly compartmentalized biochemical signalling reactions. Genetically encoded fluorescent biosensors enable the spatial and temporal monitoring of signalling events in live cells. Temporal and spatiotemporal computational models can be used to interpret biosensor experiments in complex biochemical networks and to explore hypotheses that are difficult to test experimentally. In this review, we first provide brief discussions of the experimental toolkit of fluorescent biosensors as well as computational basics with a focus on temporal and spatiotemporal deterministic models. We then describe how we used this combined approach to identify and investigate a protein kinase A (PKA) - cAMP - Ca oscillatory circuit in MIN6 β cells, a mouse pancreatic β cell system. We describe the application of this combined approach to interrogate how this oscillatory circuit is differentially regulated in a nano-compartment formed at the plasma membrane by the scaffolding protein A kinase anchoring protein 79/150. We leveraged both temporal and spatiotemporal deterministic models to identify the key regulators of this oscillatory circuit, which we confirmed with further experiments. The powerful approach of combining live-cell biosensor imaging with quantitative modelling, as discussed here, should find widespread use in the investigation of spatiotemporal regulation of cell signalling.
细胞通过将信息编码在独特分隔的生化信号反应中，对多样化的环境线索执行特定的反应。遗传编码的荧光生物传感器能够在活细胞中对信号事件进行时空监测。时间和时空计算模型可用于解释复杂生化网络中的生物传感器实验，并探索难以通过实验测试的假设。在这篇综述中，我们首先简要讨论了荧光生物传感器的实验工具包以及以时间和时空确定性模型为重点的计算基础。然后，我们描述了如何使用这种组合方法来识别和研究 MIN6 β 细胞（一种小鼠胰腺β细胞系统）中的蛋白激酶 A (PKA) - cAMP - Ca 振荡回路。我们描述了将这种组合方法应用于探究支架蛋白激酶锚定蛋白 79/150 形成的质膜纳米区室中如何对这种振荡回路进行差异调节。我们利用时间和时空确定性模型来识别这个振荡回路的关键调节因子，并用进一步的实验进行了验证。正如这里所讨论的，将活细胞生物传感器成像与定量建模相结合的强大方法，应该会在细胞信号时空调节的研究中得到广泛应用。
