活细胞中微管聚合动力学分析。
Analysis of microtubule polymerization dynamics in live cells.
作者信息
Gierke Sarah, Kumar Praveen, Wittmann Torsten
机构信息
Department of Cell & Tissue Biology, University of California, San Francisco, California 94143-0512, USA.
出版信息
Methods Cell Biol. 2010;97:15-33. doi: 10.1016/S0091-679X(10)97002-7.
Abstract
The spatiotemporal regulation of intracellular microtubule polymerization dynamics, by numerous microtubule-associated proteins and other mechanisms, is central to many cell processes. Here, we give an overview and practical guide on how to acquire and analyze time-lapse sequences of dynamic microtubules in live cells by either fluorescently labeling entire microtubules or by utilizing proteins that specifically associate only with growing microtubule ends and summarize the strengths and weaknesses of different approaches. We give practical recommendations for imaging conditions, and discuss important limitations of such analysis that are dictated by the maximum achievable spatial and temporal sampling frequencies.
摘要
细胞内微管聚合动力学的时空调节，通过众多微管相关蛋白和其他机制，是许多细胞过程的核心。在这里，我们概述并提供实用指南，介绍如何通过荧光标记整个微管或利用仅与生长中的微管末端特异性结合的蛋白质来获取和分析活细胞中动态微管的延时序列，并总结不同方法的优缺点。我们给出成像条件的实用建议，并讨论这种分析的重要局限性，这些局限性由可实现的最大空间和时间采样频率决定。
相似文献
1
Analysis of microtubule polymerization dynamics in live cells.活细胞中微管聚合动力学分析。
Methods Cell Biol. 2010;97:15-33. doi: 10.1016/S0091-679X(10)97002-7.
2
Live cell-imaging techniques for analyses of microtubules in Dictyostelium.用于分析盘基网柄菌微管的活细胞成像技术。
Methods Cell Biol. 2010;97:341-57. doi: 10.1016/S0091-679X(10)97018-0.
3
Determination of microtubule dynamic instability in living cells.活细胞中微管动态不稳定性的测定。
Methods Cell Biol. 2010;97:1-14. doi: 10.1016/S0091-679X(10)97001-5.
4
Microtubule dynamics in dendritic spines.树突棘中的微管动力学
Methods Cell Biol. 2010;97:111-32. doi: 10.1016/S0091-679X(10)97007-6.
5
Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins.通过TOG结构域蛋白重建动态微管聚合调控
Methods Enzymol. 2014;540:131-48. doi: 10.1016/B978-0-12-397924-7.00008-X.
6
Microtubule dynamics in plant cells.植物细胞中的微管动力学
Methods Cell Biol. 2010;97:373-400. doi: 10.1016/S0091-679X(10)97020-9.
7
Structure and dynamics of the kinesin-microtubule interaction revealed by fluorescence polarization microscopy.荧光偏振显微镜揭示的驱动蛋白与微管相互作用的结构和动力学
Methods Cell Biol. 2010;95:505-19. doi: 10.1016/S0091-679X(10)95025-5.
8
Analysis of microtubule dynamic instability using a plus-end growth marker.使用微管正极生长标记物分析微管动态不稳定性。
Nat Methods. 2010 Sep;7(9):761-8. doi: 10.1038/nmeth.1493. Epub 2010 Aug 22.
9
Studying plus-end tracking at single molecule resolution using TIRF microscopy.使用全内反射荧光显微镜在单分子分辨率下研究正端追踪。
Methods Cell Biol. 2010;95:543-54. doi: 10.1016/S0091-679X(10)95027-9.
10
Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy.微管动力学在体外重建并通过单分子荧光显微镜成像。
Methods Cell Biol. 2010;95:221-45. doi: 10.1016/S0091-679X(10)95013-9.
引用本文的文献
1
Anti-Breast Cancer Properties and In Vivo Safety Profile of a Bis-Carbazole Derivative.一种双咔唑衍生物的抗乳腺癌特性及体内安全性概况
Pharmaceutics. 2025 Mar 25;17(4):415. doi: 10.3390/pharmaceutics17040415.
2
Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant.生长锥的前进需要 EB1，这是通过基因组替换带有光敏感变体来揭示的。
Elife. 2023 Jan 30;12:e84143. doi: 10.7554/eLife.84143.
3
Retromer oligomerization drives SNX-BAR coat assembly and membrane constriction.Retromer 寡聚化驱动 SNX-BAR 外套组装和膜收缩。
EMBO J. 2023 Jan 16;42(2):e112287. doi: 10.15252/embj.2022112287.
4
Nanotopography modulates cytoskeletal organization and dynamics during T cell activation.纳米形貌在 T 细胞活化过程中调节细胞骨架的组织和动态。
Mol Biol Cell. 2022 Sep 1;33(10):ar88. doi: 10.1091/mbc.E21-12-0601. Epub 2022 Jul 13.
5
Functional Investigation of TUBB4A Variants Associated with Different Clinical Phenotypes.与不同临床表型相关的 TUBB4A 变异体的功能研究。
Mol Neurobiol. 2022 Aug;59(8):5056-5069. doi: 10.1007/s12035-022-02900-9. Epub 2022 Jun 6.
6
Quantification of microtubule stutters: dynamic instability behaviors that are strongly associated with catastrophe.微管猝发的量化：与崩溃密切相关的动态不稳定性行为。
Mol Biol Cell. 2022 Mar 1;33(3):ar22. doi: 10.1091/mbc.E20-06-0348. Epub 2022 Feb 2.
7
Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells.甲状腺上皮细胞纤毛中追踪胺相关受体 1 的转运。
Cells. 2021 Jun 16;10(6):1518. doi: 10.3390/cells10061518.
8
Multiple Molecular Mechanisms to Overcome Multidrug Resistance in Cancer by Natural Secondary Metabolites.天然次生代谢产物克服癌症多药耐药性的多种分子机制
Front Pharmacol. 2021 May 21;12:658513. doi: 10.3389/fphar.2021.658513. eCollection 2021.
9
Genetically encoded live-cell sensor for tyrosinated microtubules.用于酪氨酸化微管的基因编码活细胞传感器。
J Cell Biol. 2020 Oct 5;219(10). doi: 10.1083/jcb.201912107.
10
Beta3-Tubulin is Critical for Microtubule Dynamics, Cell Cycle Regulation, and Spontaneous Release of Microvesicles in Human Malignant Melanoma Cells (A375).β3-微管蛋白对于微管动力学、细胞周期调控以及人恶性黑色素瘤细胞（A375）中微囊泡的自发释放至关重要。
Int J Mol Sci. 2020 Feb 28;21(5):1656. doi: 10.3390/ijms21051656.
本文引用的文献
1
plusTipTracker: Quantitative image analysis software for the measurement of microtubule dynamics.plusTipTracker：用于测量微管动力学的定量图像分析软件。
J Struct Biol. 2011 Nov;176(2):168-84. doi: 10.1016/j.jsb.2011.07.009. Epub 2011 Jul 29.
2
Analysis of microtubule dynamic instability using a plus-end growth marker.使用微管正极生长标记物分析微管动态不稳定性。
Nat Methods. 2010 Sep;7(9):761-8. doi: 10.1038/nmeth.1493. Epub 2010 Aug 22.
3
Cell culture medium affects GFP photostability: a solution.细胞培养基会影响绿色荧光蛋白的光稳定性：一种解决方案。
Nat Methods. 2009 Dec;6(12):859-60. doi: 10.1038/nmeth1209-859.
4
Regulation of microtubule dynamic instability.微管动态不稳定性的调控
Biochem Soc Trans. 2009 Oct;37(Pt 5):1007-13. doi: 10.1042/BST0371007.
5
Effect of GFP tags on the localization of EB1 and EB1 fragments in vivo.GFP 标签对 EB1 和 EB1 片段在体内定位的影响。
Cytoskeleton (Hoboken). 2010 Jan;67(1):1-12. doi: 10.1002/cm.20409.
6
Structural plasticity in actin and tubulin polymer dynamics.肌动蛋白和微管蛋白聚合动力学中的结构可塑性
Science. 2009 Aug 21;325(5943):960-3. doi: 10.1126/science.1168823.
7
An EB1-binding motif acts as a microtubule tip localization signal.一个EB1结合基序作为微管尖端定位信号。
Cell. 2009 Jul 23;138(2):366-76. doi: 10.1016/j.cell.2009.04.065.
8
GSK3beta phosphorylation modulates CLASP-microtubule association and lamella microtubule attachment.糖原合成酶激酶3β磷酸化调节CLASP与微管的结合以及片状伪足微管附着。
J Cell Biol. 2009 Mar 23;184(6):895-908. doi: 10.1083/jcb.200901042. Epub 2009 Mar 16.
9
Mammalian end binding proteins control persistent microtubule growth.哺乳动物末端结合蛋白控制微管的持续生长。
J Cell Biol. 2009 Mar 9;184(5):691-706. doi: 10.1083/jcb.200807179. Epub 2009 Mar 2.
10
CLIP-170 tracks growing microtubule ends by dynamically recognizing composite EB1/tubulin-binding sites.CLIP-170通过动态识别复合EB1/微管蛋白结合位点来追踪生长中的微管末端。
J Cell Biol. 2008 Dec 29;183(7):1223-33. doi: 10.1083/jcb.200809190. Epub 2008 Dec 22.
Zotero 插件