scholarly journals Initiation and disassembly of filopodia tip complexes containing VASP and lamellipodin

2020 ◽  
Vol 31 (18) ◽  
pp. 2021-2034 ◽  
Author(s):  
Karen W. Cheng ◽  
R. Dyche Mullins
Keyword(s):  
Actin Filaments ◽  
Leading Edge ◽  
Live Cell ◽  
Size Dependent ◽  
Live Cell Microscopy ◽  
Cell Microscopy

Clustering of the actin polymerase VASP (vasodilator-stimulated phosphoprotein) reorganizes leading-edge actin filaments into filopodia bundles in crawling cells. Here, we use live-cell microscopy to image the earliest events in VASP clustering and find that initiation depends on interactions with lamellipodin and barbed ends, while disassembly is driven by size-dependent instability.

scholarly journals Cooperative 4Pi Excitation and Detection Yields Sevenfold Sharper Optical Sections in Live-Cell Microscopy

2004 ◽  
Vol 87 (6) ◽  
pp. 4146-4152 ◽  
Author(s):  
Hilmar Gugel ◽  
Jörg Bewersdorf ◽  
Stefan Jakobs ◽  
Johann Engelhardt ◽  
Rafael Storz ◽  
...  
Keyword(s):  
Live Cell ◽  
Live Cell Microscopy ◽  
Cell Microscopy

scholarly journals Live-cell microscopy - tips and tools

2009 ◽  
Vol 122 (6) ◽  
pp. 753-767 ◽  
Author(s):  
M. M. Frigault ◽  
J. Lacoste ◽  
J. L. Swift ◽  
C. M. Brown
Keyword(s):  
Live Cell ◽  
Live Cell Microscopy ◽  
Cell Microscopy

scholarly journals Improved Chemical-Genetic Fluorescent Markers for Live Cell Microscopy

Biochemistry ◽  
2018 ◽  
Vol 57 (39) ◽  
pp. 5648-5653 ◽  
Author(s):  
Alison G. Tebo ◽  
Frederico M. Pimenta ◽  
Yu Zhang ◽  
Arnaud Gautier
Keyword(s):  
Live Cell ◽  
Chemical Genetic ◽  
Fluorescent Markers ◽  
Live Cell Microscopy ◽  
Cell Microscopy

Dynamics and Traffic for Transfecting Exogenous MicroRNA as Studied by Live-Cell Microscopy

2021 ◽  
Vol 17 (8) ◽  
pp. 1647-1653
Author(s):  
Ke Yang ◽  
Yuanyuan Wang ◽  
Bo Sun ◽  
Tian Tian ◽  
Zhu Dai ◽  
...  
Keyword(s):  
Dynamic Properties ◽  
Mean Square Displacement ◽  
Small Region ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Calculation Results ◽  
Long Time ◽  
Live Cell Microscopy ◽  
Cell Microscopy

MicroRNA (miRNA) has emerged as an important gene-regulator that shows great potential in gene therapy because of its unique roles in gene-regulation. However, the knowledge on their function and transportation in vivo is still lacking, and there are limited obvious evidences to define intracellular transportation of miRNA. In this study, the dynamics of exogenous miR-21 transfected into HeLa cells was traced by live-cell microscopy. Their transportation at key time points was recorded and dynamic properties were analyzed by single particle tracking (SPT) and mean square displacement (MSD) calculation. Results showed that the exogenous miRNAs bounded to cells quickly and went through lysosome into cytosol, where they were subsequently recruited into p-body. They finally were degraded, otherwise went back to cytosol in some way. Long time observation and analysis of motion mode showed that the miRNAs were confined in a small region and their motion modes were flexible in different intracellular microenvironment after entering the cells.

scholarly journals Investigating the Role of F-Actin in Human Immunodeficiency Virus Assembly by Live-Cell Microscopy

2014 ◽  
Vol 88 (14) ◽  
pp. 7904-7914 ◽  
Author(s):  
Sheikh Abdul Rahman ◽  
Peter Koch ◽  
Julian Weichsel ◽  
William J. Godinez ◽  
Ulrich Schwarz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Actin Filaments ◽  
Virus Assembly ◽  
Live Cell ◽  
Viral Budding ◽  
Immunodeficiency Virus ◽  
Live Cell Microscopy ◽  
Gag Assembly ◽  
Hiv 1

ABSTRACTHuman immunodeficiency virus type 1 (HIV-1) particles assemble at the plasma membrane, which is lined by a dense network of filamentous actin (F-actin). Large amounts of actin have been detected in HIV-1 virions, proposed to be incorporated by interactions with the nucleocapsid domain of the viral polyprotein Gag. Previous studies addressing the role of F-actin in HIV-1 particle formation using F-actin-interfering drugs did not yield consistent results. Filamentous structures pointing toward nascent HIV-1 budding sites, detected by cryo-electron tomography and atomic force microscopy, prompted us to revisit the role of F-actin in HIV-1 assembly by live-cell microscopy. HeLa cells coexpressing HIV-1 carrying fluorescently labeled Gag and a labeled F-actin-binding peptide were imaged by live-cell total internal reflection fluorescence microscopy (TIR-FM). Computational analysis of image series did not reveal characteristic patterns of F-actin in the vicinity of viral budding sites. Furthermore, no transient recruitment of F-actin during bud formation was detected by monitoring fluorescence intensity changes at nascent HIV-1 assembly sites. The chosen approach allowed us to measure the effect of F-actin-interfering drugs on the assembly of individual virions in parallel with monitoring changes in the F-actin network of the respective cell. Treatment of cells with latrunculin did not affect the efficiency and dynamics of Gag assembly under conditions resulting in the disruption of F-actin filaments. Normal assembly rates were also observed upon transient stabilization of F-actin by short-term treatment with jasplakinolide. Taken together, these findings indicate that actin filament dynamics are dispensable for HIV-1 Gag assembly at the plasma membrane of HeLa cells.IMPORTANCEHIV-1 particles assemble at the plasma membrane of virus-producing cells. This membrane is lined by a dense network of actin filaments that might either present a physical obstacle to the formation of virus particles or generate force promoting the assembly process. Drug-mediated interference with the actin cytoskeleton showed different results for the formation of retroviral particles in different studies, likely due to general effects on the cell upon prolonged drug treatment. Here, we characterized the effect of actin-interfering compounds on the HIV-1 assembly process by direct observation of virus formation in live cells, which allowed us to measure assembly rate constants directly upon drug addition. Virus assembly proceeded with normal rates when actin filaments were either disrupted or stabilized. Taken together with the absence of characteristic actin filament patterns at viral budding sites in our analyses, this indicates that the actin network is dispensable for HIV-1 assembly.

Structured illumination for live cell microscopy

2018 ◽  
Author(s):  
Herbert Schneckenburger ◽  
Verena Richter ◽  
Michael Wagner ◽  
Mathis Piper
Keyword(s):  
Live Cell ◽  
Structured Illumination ◽  
Live Cell Microscopy ◽  
Cell Microscopy
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