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PLoS One. 2015 Jan 28;10(1):e0115639. doi: 10.1371/journal.pone.0115639. eCollection 2015.

In vivo imaging and characterization of actin microridges.

Author information

1
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, United States of America; Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, WI 53706, United States of America.
2
Department of Internal Medicine, Rush University Medical Center, Chicago, IL 60612, United States of America.
3
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, United States of America.
4
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, United States of America; Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53706, United States of America.

Abstract

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.

PMID:
25629723
PMCID:
PMC4309568
DOI:
10.1371/journal.pone.0115639
[Indexed for MEDLINE]
Free PMC Article

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