scholarly journals Interactions of WASp, myosin-I, and verprolin with Arp2/3 complex during actin patch assembly in fission yeast

2005 ◽  
Vol 170 (4) ◽  
pp. 637-648 ◽  
Author(s):  
Vladimir Sirotkin ◽  
Christopher C. Beltzner ◽  
Jean-Baptiste Marchand ◽  
Thomas D. Pollard
Keyword(s):  
Fission Yeast ◽  
Live Cell Imaging ◽  
Biochemical Analysis ◽  
Cell Imaging ◽  
Activator Proteins ◽  
Myosin I ◽  
Class 1 ◽  
Dynamic Structures ◽  
Actin Patch

Yeast actin patches are dynamic structures that form at the sites of cell growth and are thought to play a role in endocytosis. We used biochemical analysis and live cell imaging to investigate actin patch assembly in fission yeast Schizosaccharomyces pombe. Patch assembly proceeds via two parallel pathways: one dependent on WASp Wsp1p and verprolin Vrp1p converges with another dependent on class 1 myosin Myo1p to activate the actin-related protein 2/3 (Arp2/3) complex. Wsp1p activates Arp2/3 complex via a conventional mechanism, resulting in branched filaments. Myo1p is a weaker Arp2/3 complex activator that makes unstable branches and is enhanced by verprolin. During patch assembly in vivo, Wsp1p and Vrp1p arrive first independent of Myo1p. Arp2/3 complex associates with nascent activator patches over 6–9 s while remaining stationary. After reaching a maximum concentration, Arp2/3 complex patches move centripetally as activator proteins dissociate. Genetic dependencies of patch formation suggest that patch formation involves cross talk between Myo1p and Wsp1p/Vrp1p pathways.

scholarly journals Roles for tubulin recruitment and self-organization by TOG domain arrays in Microtubule plus-end tracking and polymerase

2018 ◽  
Author(s):  
Brian Cook ◽  
Fred Chang ◽  
Ignacio Flor-Parra ◽  
Jawdat Al-Bassam
Keyword(s):  
Fission Yeast ◽  
Functional Role ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Self Organization ◽  
Structural Studies ◽  
Overexpressed Gene

AbstractThe XMAP215/Stu2/Alp14 microtubule polymerases utilize Tumor Overexpressed Gene (TOG) domain arrays to accelerate microtubule plus-end polymerization. Structural studies suggest a microtubule polymerase model in which TOG arrays recruit four αβ-tubulins, forming large square assemblies; an array of TOG1 and TOG2 domains may then unfurl from the square state to polymerize two αβ-tubulins into protofilaments at microtubule ends. Here, we test this model using two biochemically characterized classes of fission yeast Alp14 mutants. Using in vitro reconstitution and in vivo live cell imaging, we show that αβ-tubulins recruited by TOG1 and TOG2 domains serve non-additive roles in microtubule plus-end tracking and polymerase activities. Alp14 mutants with inactivated square assembly interfaces have defects in processive plus-end tracking and poor microtubule polymerase, indicating a functional role for square assemblies in processive tracking. These studies provide functional insights into how TOG1 and TOG2 domain arrays recruit tubulins and promote polymerase at microtubule plus ends.

scholarly journals ADF/cofilin promotes invadopodial membrane recycling during cell invasion in vivo

2014 ◽  
Vol 204 (7) ◽  
pp. 1209-1218 ◽  
Author(s):  
Elliott J. Hagedorn ◽  
Laura C. Kelley ◽  
Kaleb M. Naegeli ◽  
Zheng Wang ◽  
Qiuyi Chi ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Cell Invasion ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Actin Dynamics ◽  
Membrane Structures ◽  
Tumor Dissemination ◽  
Anchor Cell ◽  
Dynamic Structures

Invadopodia are protrusive, F-actin–driven membrane structures that are thought to mediate basement membrane transmigration during development and tumor dissemination. An understanding of the mechanisms regulating invadopodia has been hindered by the difficulty of examining these dynamic structures in native environments. Using an RNAi screen and live-cell imaging of anchor cell (AC) invasion in Caenorhabditis elegans, we have identified UNC-60A (ADF/cofilin) as an essential regulator of invadopodia. UNC-60A localizes to AC invadopodia, and its loss resulted in a dramatic slowing of F-actin dynamics and an inability to breach basement membrane. Optical highlighting indicated that UNC-60A disassembles actin filaments at invadopodia. Surprisingly, loss of unc-60a led to the accumulation of invadopodial membrane and associated components within the endolysosomal compartment. Photobleaching experiments revealed that during normal invasion the invadopodial membrane undergoes rapid recycling through the endolysosome. Together, these results identify the invadopodial membrane as a specialized compartment whose recycling to form dynamic, functional invadopodia is dependent on localized F-actin disassembly by ADF/cofilin.

scholarly journals A Decade of Imaging Cellular Motility and Interaction Dynamics in the Immune System

Science ◽  
2012 ◽  
Vol 336 (6089) ◽  
pp. 1676-1681 ◽  
Author(s):  
Ronald N. Germain ◽  
Ellen A. Robey ◽  
Michael D. Cahalan
Keyword(s):  
Lymph Nodes ◽  
Live Cell Imaging ◽  
Quantitative Measurement ◽  
Cell Imaging ◽  
Plasma Cells ◽  
Effector T Cells ◽  
Cellular Interactions ◽  
Cellular Motility ◽  
Response Dynamics

To mount an immune response, lymphocytes must recirculate between the blood and lymph nodes, recognize antigens upon contact with specialized presenting cells, proliferate to expand a small number of clonally relevant lymphocytes, differentiate to antibody-producing plasma cells or effector T cells, exit from lymph nodes, migrate to tissues, and engage in host-protective activities. All of these processes involve motility and cellular interactions—events that were hidden from view until recently. Introduced to immunology by three papers in this journal in 2002, in vivo live-cell imaging studies are revealing the behavior of cells mediating adaptive and innate immunity in diverse tissue environments, providing quantitative measurement of cellular motility, interactions, and response dynamics. Here, we review themes emerging from such studies and speculate on the future of immunoimaging.

A new visible-light-excitable ICT-CHEF-mediated fluorescence ‘turn-on’ probe for the selective detection of Cd2+ in a mixed aqueous system with live-cell imaging

2015 ◽  
Vol 44 (12) ◽  
pp. 5763-5770 ◽  
Author(s):  
Shyamaprosad Goswami ◽  
Krishnendu Aich ◽  
Sangita Das ◽  
Chitrangada Das Mukhopadhyay ◽  
Deblina Sarkar ◽  
...  
Keyword(s):  
Visible Light ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Aqueous System ◽  
Live Cell ◽  
Selective Detection ◽  
Turn On ◽  
Fluorescence Turn On

A new quinoline based sensor was developed and applied for the selective detection of Cd2+ both in vitro and in vivo.

scholarly journals Regeneration of the neurogliovascular unit visualized in vivo by transcranial live-cell imaging

2020 ◽  
Vol 343 ◽  
pp. 108808 ◽  
Author(s):  
Margarita Arango-Lievano ◽  
Yann Dromard ◽  
Pierre Fontanaud ◽  
Chrystel Lafont ◽  
Patrice Mollard ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell

An aggregation-induced phosphorescence probe for calcium ion-specific detection and live-cell imaging in Arabidopsis thaliana

2019 ◽  
Vol 55 (33) ◽  
pp. 4841-4844 ◽  
Author(s):  
Guilin Chen ◽  
Zaicai Zhou ◽  
Hui Feng ◽  
Chenyan Zhang ◽  
Yifan Wang ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Calcium Ion ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Molecular Probe ◽  
Specific Detection ◽  
Phosphorescence Probe

A molecular probe with aggregation-induced phosphorescence (AIP) properties for calcium ion-specific detection and imaging in vivo was designed.

scholarly journals Detection and Characterization of Protein Interactions In Vivo by a Simple Live-Cell Imaging Method

PLoS ONE ◽  
2013 ◽  
Vol 8 (5) ◽  
pp. e62195 ◽  
Author(s):  
Oriol Gallego ◽  
Tanja Specht ◽  
Thorsten Brach ◽  
Arun Kumar ◽  
Anne-Claude Gavin ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Imaging Method

scholarly journals Live Cell Imaging of Butterfly Pupal and Larval Wings In Vivo

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0128332 ◽  
Author(s):  
Yoshikazu Ohno ◽  
Joji M. Otaki
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell
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