Biological Physics, Department of Physics and Astronomy, Schuster Building, The University of Manchester, Oxford Road, Manchester, M13 9PL, UK.
Division of Molecular and Cellular Function, School of Biological Sciences, Michael Smith Building, The University of Manchester, Dover Street, Manchester, M13 9PT, UK.
Sci Rep. 2021 Aug 10;11(1):16230. doi: 10.1038/s41598-021-94901-2.
The endoplasmic reticulum (ER) is a eukaryotic subcellular organelle composed of tubules and sheet-like areas of membrane connected at junctions. The tubule network is highly dynamic and undergoes rapid and continual rearrangement. There are currently few tools to evaluate network organisation and dynamics. We quantified ER network organisation in Vero and MRC5 cells, and developed an analysis workflow for dynamics of established tubules in live cells. The persistence length, tubule length, junction coordination number and angles of the network were quantified. Hallmarks of imbalances in ER tension, indications of interactions with microtubules and other subcellular organelles, and active dynamics were observed. Clear differences in dynamic behaviour were observed for established tubules at different positions within the cell using itemset mining. We found that tubules with activity-driven fluctuations were more likely to be located away from the cell periphery and a population of peripheral tubules with no signs of active motion was found.
内质网(ER)是真核细胞的亚细胞细胞器,由小管和膜的片状区域组成,在连接处连接。小管网络具有高度动态性,并经历快速和连续的重新排列。目前很少有工具来评估网络组织和动态。我们在 Vero 和 MRC5 细胞中量化了 ER 网络组织,并为活细胞中已建立的小管的动力学开发了一种分析工作流程。网络的持久长度、小管长度、连接协调数和角度都进行了量化。观察到 ER 张力失衡的特征、与微管和其他亚细胞细胞器相互作用的迹象以及活跃的动力学。使用项集挖掘观察到细胞内不同位置的已建立小管的明显不同的动态行为。我们发现,具有活性驱动波动的小管更有可能位于远离细胞边缘的位置,并且发现了一群没有活性运动迹象的外围小管。