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In Vivo Imaging and Characterization of Actin Microridges

Actin microridges form labyrinth like patterns on superficial epithelial cells across animal species. This highly organized assembly has been implicated in mucus retention and in the mechanical structure of mucosal surfaces, however the mechanisms that regulate actin microridges remain largely unkno...

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Autores principales: Lam, Pui-ying, Mangos, Steve, Green, Julie M., Reiser, Jochen, Huttenlocher, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309568/
https://www.ncbi.nlm.nih.gov/pubmed/25629723
http://dx.doi.org/10.1371/journal.pone.0115639
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author Lam, Pui-ying
Mangos, Steve
Green, Julie M.
Reiser, Jochen
Huttenlocher, Anna
author_facet Lam, Pui-ying
Mangos, Steve
Green, Julie M.
Reiser, Jochen
Huttenlocher, Anna
author_sort Lam, Pui-ying
collection PubMed
description Actin microridges form labyrinth like patterns on superficial epithelial cells across animal species. This highly organized assembly has been implicated in mucus retention and in the mechanical structure of mucosal surfaces, however the mechanisms that regulate actin microridges remain largely unknown. Here we characterize the composition and dynamics of actin microridges on the surface of zebrafish larvae using live imaging. Microridges contain phospho-tyrosine, cortactin and VASP, but not focal adhesion kinase. Time-lapse imaging reveals dynamic changes in the length and branching of microridges in intact animals. Transient perturbation of the microridge pattern occurs before cell division with rapid re-assembly during and after cytokinesis. Microridge assembly is maintained with constitutive activation of Rho or inhibition of myosin II activity. However, expression of dominant negative RhoA or Rac alters microridge organization, with an increase in distance between microridges. Latrunculin A treatment and photoconversion experiments suggest that the F-actin filaments are actively treadmilling in microridges. Accordingly, inhibition of Arp2/3 or PI3K signaling impairs microridge structure and length. Taken together, actin microridges in zebrafish represent a tractable in vivo model to probe pattern formation and dissect Arp2/3-mediated actin dynamics in vivo.
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spelling pubmed-43095682015-02-06 In Vivo Imaging and Characterization of Actin Microridges Lam, Pui-ying Mangos, Steve Green, Julie M. Reiser, Jochen Huttenlocher, Anna PLoS One Research Article Actin microridges form labyrinth like patterns on superficial epithelial cells across animal species. This highly organized assembly has been implicated in mucus retention and in the mechanical structure of mucosal surfaces, however the mechanisms that regulate actin microridges remain largely unknown. Here we characterize the composition and dynamics of actin microridges on the surface of zebrafish larvae using live imaging. Microridges contain phospho-tyrosine, cortactin and VASP, but not focal adhesion kinase. Time-lapse imaging reveals dynamic changes in the length and branching of microridges in intact animals. Transient perturbation of the microridge pattern occurs before cell division with rapid re-assembly during and after cytokinesis. Microridge assembly is maintained with constitutive activation of Rho or inhibition of myosin II activity. However, expression of dominant negative RhoA or Rac alters microridge organization, with an increase in distance between microridges. Latrunculin A treatment and photoconversion experiments suggest that the F-actin filaments are actively treadmilling in microridges. Accordingly, inhibition of Arp2/3 or PI3K signaling impairs microridge structure and length. Taken together, actin microridges in zebrafish represent a tractable in vivo model to probe pattern formation and dissect Arp2/3-mediated actin dynamics in vivo. Public Library of Science 2015-01-28 /pmc/articles/PMC4309568/ /pubmed/25629723 http://dx.doi.org/10.1371/journal.pone.0115639 Text en © 2015 Lam et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Lam, Pui-ying
Mangos, Steve
Green, Julie M.
Reiser, Jochen
Huttenlocher, Anna
In Vivo Imaging and Characterization of Actin Microridges
title In Vivo Imaging and Characterization of Actin Microridges
title_full In Vivo Imaging and Characterization of Actin Microridges
title_fullStr In Vivo Imaging and Characterization of Actin Microridges
title_full_unstemmed In Vivo Imaging and Characterization of Actin Microridges
title_short In Vivo Imaging and Characterization of Actin Microridges
title_sort in vivo imaging and characterization of actin microridges
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309568/
https://www.ncbi.nlm.nih.gov/pubmed/25629723
http://dx.doi.org/10.1371/journal.pone.0115639
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