Comparative analysis of tools for live cell imaging of actin network architecture.

Bioarchitecture
Brittany J BelinR Dyche Mullins

Abstract

Fluorescent derivatives of actin and actin-binding domains are powerful tools for studying actin filament architecture and dynamics in live cells. Growing evidence, however, indicates that these probes are biased, and their cellular distribution does not accurately reflect that of the cytoskeleton. To understand the strengths and weaknesses of commonly used live-cell probes--fluorescent protein fusions of actin, Lifeact, F-tractin, and actin-binding domains from utrophin--we compared their distributions in cells derived from various model organisms. We focused on five actin networks: the peripheral cortex, lamellipodial and lamellar networks, filopodial bundles, and stress fibers. Using phalloidin as a standard, we identified consistent biases in the distribution of each probe. The localization of F-tractin is the most similar to that of phalloidin but induces organism-specific changes in cell morphology. Both Lifeact and GFP-actin concentrate in lamellipodial actin networks but are excluded from lamellar networks and filopodia. In contrast, the full utrophin actin-binding domain (Utr261) binds filaments of the lamellum but only weakly localizes to lamellipodia, while a shorter variant (Utr230) is restricted to the most stable ...Continue Reading

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Citations

Feb 6, 2016·Histochemistry and Cell Biology·Ildikó KristóPéter Vilmos
Nov 10, 2015·Experimental Cell Research·Aleksandra SimiczyjewDorota Nowak
Jan 23, 2016·Journal of Biomechanics·Kristina SliogeryteMartin M Knight
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Jun 23, 2020·The EMBO Journal·Archana KumariMinhajuddin Sirajuddin
Sep 19, 2020·Proceedings of the National Academy of Sciences of the United States of America·Hawa Racine ThiamClare M Waterman
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Jan 14, 2017·Journal of Cell Science·Michael MelakRobert Grosse
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Methods Mentioned

BETA
fluorescence recovery after photobleaching
transfection
fluorescence recovery
transfections

Software Mentioned

ImageJ
R
CurveExpertPro

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