Actin visualization at a glance

被引:172
作者
Melak, Michael [1 ]
Plessner, Matthias [1 ]
Grosse, Robert [1 ]
机构
[1] Univ Marburg, Biochem Pharmacol Ctr, Inst Pharmacol, Karl von Frisch Str 1, D-35043 Marburg, Germany
关键词
Actin dynamics; Actin probes; Live-cell imaging; Nuclear actin; F-ACTIN; LIVE CELLS; IN-VIVO; CYTOSKELETAL DYNAMICS; NONMUSCLE CELLS; ARP2/3; COMPLEX; FUSION PROTEIN; LIFEACT; BINDING; FILAMENTS;
D O I
10.1242/jcs.189068
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Actin functions in a multitude of cellular processes owing to its ability to polymerize into filaments, which can be further organized into higher-order structures by an array of actin-binding and regulatory proteins. Therefore, research on actin and actin-related functions relies on the visualization of actin structures without interfering with the cycles of actin polymerization and depolymerization that underlie cellular actin dynamics. In this Cell Science at a Glance and the accompanying poster, we briefly evaluate the different techniques and approaches currently applied to analyze and visualize cellular actin structures, including in the nuclear compartment. Referring to the gold standard F-actin marker phalloidin to stain actin in fixed samples and tissues, we highlight methods for visualization of actin in living cells, which mostly apply the principle of genetically fusing fluorescent proteins to different actin-binding domains, such as LifeAct, utrophin and F-tractin, as well as anti-actin-nanobody technology. In addition, the compound SiR-actin and the expression of GFP-actin are also applicable for various types of live-cell analyses. Overall, the visualization of actin within a physiological context requires a careful choice of method, as well as a tight control of the amount or the expression level of a given detection probe in order to minimize its influence on endogenous actin dynamics.
引用
收藏
页码:525 / 530
页数:6
相关论文
共 77 条
[1]   Nuclear Actin Network Assembly by Formins Regulates the SRF Coactivator MAL [J].
Baarlink, Christian ;
Wang, Haicui ;
Grosse, Robert .
SCIENCE, 2013, 340 (6134) :864-867
[2]  
Ballestrem C, 1998, J CELL SCI, V111, P1649
[3]  
Belin BJ, 2015, ELIFE, V4, DOI [10.7554/eLife.07735, 10.7554/eLife.11935]
[4]  
Belin Brittany J, 2014, Bioarchitecture, V4, P189, DOI 10.1080/19490992.2014.1047714
[5]   F-Actin Dynamics in Neurospora crassa [J].
Berepiki, Adokiye ;
Lichius, Alexander ;
Shoji, Jun-Ya ;
Tilsner, Jens ;
Read, Nick D. .
EUKARYOTIC CELL, 2010, 9 (04) :547-557
[6]  
BUBB MR, 1994, J BIOL CHEM, V269, P14869
[7]   LOV-based reporters for fluorescence imaging [J].
Buckley, Anthony M. ;
Petersen, Jan ;
Roe, Andrew J. ;
Douce, Gillian R. ;
Christie, John M. .
CURRENT OPINION IN CHEMICAL BIOLOGY, 2015, 27 :39-45
[8]   β-Actin specifically controls cell growth, migration, and the G-actin pool [J].
Bunnell, Tina M. ;
Burbach, Brandon J. ;
Shimizu, Yoji ;
Ervasti, James M. .
MOLECULAR BIOLOGY OF THE CELL, 2011, 22 (21) :4047-4058
[9]   Versatile fluorescent probes for actin filaments based on the actin-binding domain of Utrophin [J].
Burkel, Brian M. ;
von Dassow, George ;
Bement, William M. .
CELL MOTILITY AND THE CYTOSKELETON, 2007, 64 (11) :822-832
[10]   Structural Memory in the Contractile Ring Makes the Duration of Cytokinesis Independent of Cell Size [J].
Carvalho, Ana ;
Desai, Arshad ;
Oegema, Karen .
CELL, 2009, 137 (05) :926-937