使用鬼笔环肽在……中可视化丝状肌动蛋白
Visualizing Filamentous Actin Using Phalloidin in .
作者信息
Craig Evan W, Avasthi Prachee
机构信息
Anatomy & Cell Biology, University of Kansas Medical Center, Kansas City, KS, USA.
出版信息
Bio Protoc. 2019 Jun 20;9(12). doi: 10.21769/BioProtoc.3274.
Abstract
This protocol aims to visualize the filamentous actin network in . We improved fixed-cell labeling conditions using the F-actin probe, phalloidin. We created a -optimized protocol by halving the phalloidin incubation time, electing for optimal fixation conditions, and selecting for a healthy cell population. This phalloidin protocol is quick, effective, and is the only labeling method to date that allows for reliable actin filament detection in fixed vegetative cells. This method reveals previously unidentified actin structures in and novel insights into cytoskeletal dynamics.
摘要
本方案旨在可视化……中的丝状肌动蛋白网络。我们使用F-肌动蛋白探针鬼笔环肽改进了固定细胞标记条件。我们通过将鬼笔环肽孵育时间减半、选择最佳固定条件以及挑选健康细胞群体，创建了一种优化方案。这种鬼笔环肽方案快速、有效，是迄今为止唯一一种能够在固定的营养细胞中可靠检测肌动蛋白丝的标记方法。该方法揭示了……中以前未被识别的肌动蛋白结构以及对细胞骨架动力学的新见解。
相似文献
1
Visualizing Filamentous Actin Using Phalloidin in .使用鬼笔环肽在……中可视化丝状肌动蛋白
Bio Protoc. 2019 Jun 20;9(12). doi: 10.21769/BioProtoc.3274.
2
The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins.表达常规和不同形式肌动蛋白的绿藻中难以捉摸的肌动蛋白细胞骨架。
Mol Biol Cell. 2019 Oct 15;30(22):2827-2837. doi: 10.1091/mbc.E19-03-0141. Epub 2019 Sep 18.
3
Labeling cytoskeletal F-actin with rhodamine phalloidin or fluorescein phalloidin for imaging.用罗丹明鬼笔环肽或荧光素鬼笔环肽标记细胞骨架F-肌动蛋白以进行成像。
Cold Spring Harb Protoc. 2010 May;2010(5):pdb.prot4947. doi: 10.1101/pdb.prot4947.
4
Actin filament organization in the fish keratocyte lamellipodium.鱼类角膜细胞片状伪足中的肌动蛋白丝组织
J Cell Biol. 1995 Jun;129(5):1275-86. doi: 10.1083/jcb.129.5.1275.
5
Visualization of endogenous nuclear F-actin in mouse embryos reveals abnormal actin assembly after somatic cell nuclear transfer.可视化内源核 F-actin 在小鼠胚胎中的表达揭示了体细胞核移植后肌动蛋白组装异常。
J Biochem. 2021 Apr 18;169(3):303-311. doi: 10.1093/jb/mvaa125.
6
Quantitative Superresolution Imaging of F-Actin in the Cell Body and Cytoskeletal Protrusions Using Phalloidin-Based Single-Molecule Labeling and Localization Microscopy.使用基于鬼笔环肽的单分子标记和定位显微镜对细胞体和细胞骨架突起中的F-肌动蛋白进行定量超分辨率成像。
bioRxiv. 2024 Mar 6:2024.03.04.583337. doi: 10.1101/2024.03.04.583337.
7
Actin dynamics during the cell cycle in Chlamydomonas reinhardtii.莱茵衣藻细胞周期中的肌动蛋白动力学
Cell Motil Cytoskeleton. 1992;22(2):117-26. doi: 10.1002/cm.970220205.
8
Improved visualization of F-actin in the green alga Acetabularia by microwave-accelerated fixation and simultaneous FITC-Phalloidin staining.通过微波加速固定和同时进行异硫氰酸荧光素-鬼笔环肽染色改善绿藻伞藻中F-肌动蛋白的可视化。
Histochem J. 1996 May;28(5):353-60. doi: 10.1007/BF02331398.
9
Phalloidin Staining of Actin Filaments for Visualization of Muscle Fibers in .用于可视化……中肌纤维的肌动蛋白丝的鬼笔环肽染色
Bio Protoc. 2021 Oct 5;11(19):e4183. doi: 10.21769/BioProtoc.4183.
10
Filamentous actin in Paramecium cells: mapping by phalloidin affinity labeling in vivo and in vitro.草履虫细胞中的丝状肌动蛋白：通过鬼笔环肽亲和标记在体内和体外进行定位
J Histochem Cytochem. 1986 Apr;34(4):443-54. doi: 10.1177/34.4.2419395.
引用本文的文献
1
Mitochondria dysfunction is one of the causes of diclofenac toxicity in the green alga .线粒体功能障碍是二氯芬酸对绿藻毒性的原因之一。
PeerJ. 2024 Aug 28;12:e18005. doi: 10.7717/peerj.18005. eCollection 2024.
2
The ERK activator, BCI, inhibits ciliogenesis and causes defects in motor behavior, ciliary gating, and cytoskeletal rearrangement.ERK 激活剂 BCI 抑制纤毛生成，并导致运动行为、纤毛门控和细胞骨架重排缺陷。
Life Sci Alliance. 2023 Mar 13;6(6). doi: 10.26508/lsa.202301899. Print 2023 Jun.
3
Lithium-induced ciliary lengthening sparks Arp2/3 complex-dependent endocytosis.锂诱导的纤毛伸长引发 Arp2/3 复合物依赖性内吞作用。
Mol Biol Cell. 2023 Apr 1;34(4):ar26. doi: 10.1091/mbc.E22-06-0219. Epub 2023 Feb 8.
4
Novel Cyclic Peptides from Lethal Mushrooms through a Genome-Guided Approach.通过基因组导向方法从致命蘑菇中发现的新型环肽
J Fungi (Basel). 2021 Mar 11;7(3):204. doi: 10.3390/jof7030204.
5
The Anti-Cancer Effect of Linusorb B3 from Flaxseed Oil through the Promotion of Apoptosis, Inhibition of Actin Polymerization, and Suppression of Src Activity in Glioblastoma Cells.亚麻籽油中 B3 对脑胶质瘤细胞的抗癌作用是通过促进细胞凋亡、抑制肌动蛋白聚合和抑制 Src 活性实现的。
Molecules. 2020 Dec 12;25(24):5881. doi: 10.3390/molecules25245881.
6
formin FOR1 and profilin PRF1 are optimized for acute rapid actin filament assembly.formin FOR1 和 profilin PRF1 被优化用于急性快速肌动蛋白丝组装。
Mol Biol Cell. 2019 Dec 15;30(26):3123-3135. doi: 10.1091/mbc.E19-08-0463. Epub 2019 Oct 30.
7
The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins.表达常规和不同形式肌动蛋白的绿藻中难以捉摸的肌动蛋白细胞骨架。
Mol Biol Cell. 2019 Oct 15;30(22):2827-2837. doi: 10.1091/mbc.E19-03-0141. Epub 2019 Sep 18.
本文引用的文献
1
Lifeact-GFP alters F-actin organization, cellular morphology and biophysical behaviour.Lifeact-GFP 改变 F-肌动蛋白的组织、细胞形态和生物物理行为。
Sci Rep. 2019 Mar 1;9(1):3241. doi: 10.1038/s41598-019-40092-w.
2
Avoiding artefacts when counting polymerized actin in live cells with LifeAct fused to fluorescent proteins.当使用与荧光蛋白融合的LifeAct对活细胞中的聚合肌动蛋白进行计数时避免假象。
Nat Cell Biol. 2016 Jun;18(6):676-83. doi: 10.1038/ncb3351. Epub 2016 May 9.
3
Evidence That an Unconventional Actin Can Provide Essential F-Actin Function and That a Surveillance System Monitors F-Actin Integrity in Chlamydomonas.衣藻中一种非常规肌动蛋白可提供必需的F-肌动蛋白功能以及一种监测系统可监控F-肌动蛋白完整性的证据。
Genetics. 2016 Mar;202(3):977-96. doi: 10.1534/genetics.115.184663. Epub 2015 Dec 29.
4
Actin is required for IFT regulation in Chlamydomonas reinhardtii.肌动蛋白对于莱茵衣藻中的IFT调节是必需的。
Curr Biol. 2014 Sep 8;24(17):2025-32. doi: 10.1016/j.cub.2014.07.038. Epub 2014 Aug 21.
5
NIH Image to ImageJ: 25 years of image analysis.NIH 图像到 ImageJ：25 年的图像分析。
Nat Methods. 2012 Jul;9(7):671-5. doi: 10.1038/nmeth.2089.
6
Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas.衣藻核转化体中外源基因的表观遗传沉默
Plant Cell. 1997 Jun;9(6):925-45. doi: 10.1105/tpc.9.6.925.
7
Localization of actin in Chlamydomonas using antiactin and NBD-phallacidin.利用抗肌动蛋白和NBD-鬼笔环肽对衣藻中的肌动蛋白进行定位。
Cell Motil. 1985;5(5):415-30. doi: 10.1002/cm.970050505.
8
Actin dynamics during the cell cycle in Chlamydomonas reinhardtii.莱茵衣藻细胞周期中的肌动蛋白动力学
Cell Motil Cytoskeleton. 1992;22(2):117-26. doi: 10.1002/cm.970220205.
Zotero 插件