Institut de Biologie Moléculaire des Plantes (IBMP), CNRS, Université de Strasbourg, Strasbourg, France.
Université Aix Marseille, CEA, CNRS, BIAM, UMR7265, Saint-Paul-lez-Durance, France.
Methods Mol Biol. 2025;2873:223-245. doi: 10.1007/978-1-0716-4228-3_13.
Nuclear dynamics refers to global/local changes in the molecular and spatial organization of genomic DNA that can occur during development or in response to environmental stress signals and eventually impact genomic functions. In plants, nuclear dynamics relies notably on the connection of the nucleus with the cytoskeleton during development. It orchestrates genomic functions in response to developmental and environmental cues. This is particularly true in the plant root system, which is constantly exposed to a wide range of internal and external stimuli. Currently, studying nuclear dynamics in a growing root is challenging due to limitations regarding real-time imaging for quantitative analyses under controlled conditions. Microfluidic systems for plant cell studies are valuable analytical tools that provide precise control of culture conditions together with live-imaging capabilities at high temporal and spatial resolutions. Herein, we describe a microfluidic platform to unravel dynamically and noninvasively nuclear organization in the seedling root system exposed to various treatments. As exemplified here, our microfluidic platform can be conveniently used for real-time microscopy imaging and quantitative analysis of fine nuclear morphological changes upon modifying cytoskeleton dynamics. Importantly, our system can be applied to a wide variety of microscopic means including high-resolution microscopy to investigate diverse subcellular compartments or nuclear domains in Arabidopsis thaliana roots.
核动态是指基因组 DNA 的分子和空间组织在发育过程中或对外界环境压力信号的反应中发生的全局/局部变化,最终会影响基因组功能。在植物中,核动态主要依赖于发育过程中细胞核与细胞骨架的连接。它通过响应发育和环境线索来协调基因组功能。在植物根系中,这一点尤为明显,因为根系不断受到各种内部和外部刺激的影响。目前,由于在受控条件下进行实时成像进行定量分析的限制,研究生长中的根中的核动态具有挑战性。用于植物细胞研究的微流控系统是有价值的分析工具,它提供了对培养条件的精确控制以及在高时空分辨率下的实时成像能力。在这里,我们描述了一种微流控平台,用于揭示暴露于各种处理的幼苗根系中动态且非侵入性的核组织。如这里所示,我们的微流控平台可方便地用于实时显微镜成像和定量分析在改变细胞骨架动力学时精细的核形态变化。重要的是,我们的系统可应用于各种微观手段,包括高分辨率显微镜,以研究拟南芥根中的各种亚细胞区室或核域。