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Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation

Photoactivation allows one to pulse-label molecules and obtain quantitative data about their behavior. We have devised a new modeling-based analysis for photoactivatable actin experiments that simultaneously measures properties of monomeric and filamentous actin in a three-dimensional cellular envir...

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Detalles Bibliográficos
Autores principales: Kapustina, Maryna, Read, Tracy-Ann, Vitriol, Eric A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5201019/
https://www.ncbi.nlm.nih.gov/pubmed/27831495
http://dx.doi.org/10.1242/jcs.194670
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author Kapustina, Maryna
Read, Tracy-Ann
Vitriol, Eric A.
author_facet Kapustina, Maryna
Read, Tracy-Ann
Vitriol, Eric A.
author_sort Kapustina, Maryna
collection PubMed
description Photoactivation allows one to pulse-label molecules and obtain quantitative data about their behavior. We have devised a new modeling-based analysis for photoactivatable actin experiments that simultaneously measures properties of monomeric and filamentous actin in a three-dimensional cellular environment. We use this method to determine differences in the dynamic behavior of β- and γ-actin isoforms, showing that both inhabit filaments that depolymerize at equal rates but that β-actin exists in a higher monomer-to-filament ratio. We also demonstrate that cofilin (cofilin 1) equally accelerates depolymerization of filaments made from both isoforms, but is only required to maintain the β-actin monomer pool. Finally, we used modeling-based analysis to assess actin dynamics in axon-like projections of differentiating neuroblastoma cells, showing that the actin monomer concentration is significantly depleted as the axon develops. Importantly, these results would not have been obtained using traditional half-time analysis. Given that parameters of the publicly available modeling platform can be adjusted to suit the experimental system of the user, this method can easily be used to quantify actin dynamics in many different cell types and subcellular compartments.
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spelling pubmed-52010192017-01-23 Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation Kapustina, Maryna Read, Tracy-Ann Vitriol, Eric A. J Cell Sci Tools and Techniques Photoactivation allows one to pulse-label molecules and obtain quantitative data about their behavior. We have devised a new modeling-based analysis for photoactivatable actin experiments that simultaneously measures properties of monomeric and filamentous actin in a three-dimensional cellular environment. We use this method to determine differences in the dynamic behavior of β- and γ-actin isoforms, showing that both inhabit filaments that depolymerize at equal rates but that β-actin exists in a higher monomer-to-filament ratio. We also demonstrate that cofilin (cofilin 1) equally accelerates depolymerization of filaments made from both isoforms, but is only required to maintain the β-actin monomer pool. Finally, we used modeling-based analysis to assess actin dynamics in axon-like projections of differentiating neuroblastoma cells, showing that the actin monomer concentration is significantly depleted as the axon develops. Importantly, these results would not have been obtained using traditional half-time analysis. Given that parameters of the publicly available modeling platform can be adjusted to suit the experimental system of the user, this method can easily be used to quantify actin dynamics in many different cell types and subcellular compartments. The Company of Biologists Ltd 2016-12-15 /pmc/articles/PMC5201019/ /pubmed/27831495 http://dx.doi.org/10.1242/jcs.194670 Text en © 2016. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Tools and Techniques
Kapustina, Maryna
Read, Tracy-Ann
Vitriol, Eric A.
Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title_full Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title_fullStr Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title_full_unstemmed Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title_short Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
title_sort simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
topic Tools and Techniques
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5201019/
https://www.ncbi.nlm.nih.gov/pubmed/27831495
http://dx.doi.org/10.1242/jcs.194670
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