Department of Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan.
Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan.
J Cell Physiol. 2021 Jun;236(6):4681-4693. doi: 10.1002/jcp.30190. Epub 2020 Nov 26.
The ability of a single Ca ion to play an important role in cell biology is highlighted by the need for cells to form Ca signals in the dimensions of space, time, and amplitude. Thus, spatial and temporal changes in intracellular Ca concentration are important for determining cell fate. Optogenetic technology has been developed to provide more precise and targeted stimulation of cells. Here, U2OS cells overexpressing Ca translocating channelrhodopsin (CatCh) were used to mediate Ca influx through blue light illumination with various parameters, such as intensity, frequency, duty cycle, and duration. We identified that several Ca -dependent transcription factors and certain kinases can be activated by specific Ca waves. Using a wound-healing assay, we found that low-frequency Ca oscillations increased cell migration through the activation of NF-κB. This study explores the regulation of cell migration by Ca signals. Thus, we can choose optical parameters to modulate Ca waves and achieve activation of specific signaling pathways. This novel methodology can be applied to clarify related cell-signaling mechanisms in the future.
钙离子在细胞生物学中发挥重要作用的能力，突出表现在细胞需要在空间、时间和幅度维度上形成钙信号。因此，细胞内钙离子浓度的时空变化对于决定细胞命运至关重要。光遗传学技术的发展为细胞的精确和靶向刺激提供了可能。在这里，过表达钙转运通道视紫红质（CatCh）的 U2OS 细胞被用于通过不同参数（如强度、频率、占空比和持续时间）的蓝光照射来介导钙内流。我们发现，几种钙依赖性转录因子和某些激酶可以被特定的钙波激活。通过划痕实验，我们发现低频钙振荡通过激活 NF-κB 增加细胞迁移。本研究探讨了钙信号对细胞迁移的调控。因此，我们可以选择光学参数来调制钙波，并实现特定信号通路的激活。这种新的方法学可以应用于未来阐明相关的细胞信号机制。
