Max Planck Institute for Dynamics and Self-Organization, Göttingen, Germany.
School of Mathematics and Physics, University of Surrey, Guildford, United Kingdom.
Methods Mol Biol. 2024;2828:107-117. doi: 10.1007/978-1-0716-4023-4_10.
Living cells have the ability to detect electric fields and respond to them with directed migratory movements. Many proteomic approaches have been adopted in the past to identify the molecular mechanism behind this cellular phenomenon. However, how the cells sense the electric stimulus and transduce it into directed cell migration is still under discussion. Many eukaryotic cells react to applied electric stimulation, including Dictyostelium discoideum cells. We use them as model system for studying cell migration in electric fields, also known as electrotaxis. Here we report the protocols that we developed for our experiments. Our experimental outcomes helped us to characterize: (i) the memory that cells have in a varying electric field, which we defined as temporal electric persistence; and (ii) the accelerating motion of cells along their paths over the electric exposure time. We also report on the analysis of the role that conditioned medium factor (CMF), a protein secreted by cells when they begin to starve, plays in the mechanism of electric sensing. The results of this study can contribute to the understanding of the electrical sensing of cells and its transduction into directed cell migration.
活细胞具有检测电场的能力，并能通过定向迁移运动对其做出反应。过去，人们采用了许多蛋白质组学方法来确定这一细胞现象背后的分子机制。然而，细胞如何感知电刺激并将其转化为定向细胞迁移仍在讨论中。许多真核细胞对施加的电刺激有反应，包括盘基网柄菌细胞。我们将它们用作研究电场中细胞迁移的模型系统，也称为趋电性。本文报告了我们为实验开发的方案。我们的实验结果帮助我们描述了：（i）细胞在变化的电场中具有的记忆，我们将其定义为时间电持久性；（ii）细胞在暴露于电场的过程中沿其路径加速运动的情况。我们还报告了分析条件培养基因子 (CMF) 在电感应机制中的作用，CMF 是细胞在饥饿开始时分泌的一种蛋白质。这项研究的结果有助于理解细胞的电感应及其转化为定向细胞迁移。
