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微流控技术用于追踪植物根生长过程中核质界面的时空动态。

Microfluidics to Follow Spatiotemporal Dynamics at the Nucleo-Cytoplasmic Interface During Plant Root Growth.

机构信息

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.

出版信息

Methods Mol Biol. 2025;2873:223-245. doi: 10.1007/978-1-0716-4228-3_13.

DOI:10.1007/978-1-0716-4228-3_13
PMID:39576605
Abstract

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.

摘要

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。

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本文引用的文献

1
Plant nuclear envelope as a hub connecting genome organization with regulation of gene expression.植物核被膜作为连接基因组组织与基因表达调控的枢纽。
Nucleus. 2023 Dec;14(1):2178201. doi: 10.1080/19491034.2023.2178201.
2
Tools for studying the cytoskeleton during plant cell division.研究植物细胞分裂过程中细胞骨架的工具。
Trends Plant Sci. 2022 Oct;27(10):1049-1062. doi: 10.1016/j.tplants.2022.05.006. Epub 2022 Jun 3.
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Imaging the living plant cell: From probes to quantification.活体植物细胞成像:从探针到定量。
Plant Cell. 2022 Jan 20;34(1):247-272. doi: 10.1093/plcell/koab237.
4
Real-time tracking of root hair nucleus morphodynamics using a microfluidic approach.使用微流控方法实时跟踪根毛细胞核形态动力学。
Plant J. 2021 Oct;108(2):303-313. doi: 10.1111/tpj.15511. Epub 2021 Oct 10.
5
Tracking Root Interactions System (TRIS) Experiment and Quality Control.根系相互作用追踪系统(TRIS)实验与质量控制。
Bio Protoc. 2019 Apr 20;9(8):e3211. doi: 10.21769/BioProtoc.3211.
6
Tissue folding at the organ-meristem boundary results in nuclear compression and chromatin compaction.器官分生组织边界处的组织折叠导致核压缩和染色质紧缩。
Proc Natl Acad Sci U S A. 2021 Feb 23;118(8). doi: 10.1073/pnas.2017859118.
7
Chromatin Manipulation and Editing: Challenges, New Technologies and Their Use in Plants.染色质操作和编辑:挑战、新技术及其在植物中的应用。
Int J Mol Sci. 2021 Jan 6;22(2):512. doi: 10.3390/ijms22020512.
8
Subnuclear gene positioning through lamina association affects copper tolerance.通过核层关联进行亚核基因定位会影响铜耐受性。
Nat Commun. 2020 Nov 20;11(1):5914. doi: 10.1038/s41467-020-19621-z.
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Automated 3D bio-imaging analysis of nuclear organization by NucleusJ 2.0.NucleusJ 2.0 实现的细胞核三维生物成像分析自动化。
Nucleus. 2020 Dec;11(1):315-329. doi: 10.1080/19491034.2020.1845012.
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Is the plant nucleus a mechanical rheostat?植物细胞核是机械变阻器吗?
Curr Opin Plant Biol. 2020 Oct;57:155-163. doi: 10.1016/j.pbi.2020.09.001. Epub 2020 Oct 29.