Simultaneous quantification of actin monomer and filament dynamics with modeling-assisted analysis of photoactivation
Autor: | Tracy Ann Read, Maryna Kapustina, Eric A. Vitriol |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Photoactivation Light Green Fluorescent Proteins macromolecular substances Biology Filamentous actin Models Biological Protein filament 03 medical and health sciences chemistry.chemical_compound Cofilin 1 Cell Line Tumor Tools and Techniques Animals Humans Actin Depolymerization Modeling Cell Differentiation Cell Biology Cofilin Actin cytoskeleton Actins Axons 3. Good health Cell biology Actin Cytoskeleton 030104 developmental biology Monomer chemistry Axonal cytoskeleton |
Zdroj: | Journal of Cell Science |
ISSN: | 1477-9137 |
Popis: | 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. Summary: We have devised a new modeling-based analysis for photoactivatable actin experiments that accurately measures multiple parameters of actin dynamics, providing more detailed results than those obtained using traditional half-time analysis. |
Databáze: | OpenAIRE |
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