In vivo imaging and characterization of actin microridges

Pui Ying Lam, Steve Mangos, Julie M. Green, Jochen Reiser, Anna Huttenlocher

Research output: Contribution to journalArticlepeer-review

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 ofmucosal surfaces, however themechanisms that regulate actin microridges remain largely unknown. Here we characterize the composition and dynamics of actinmicroridges 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.

Original languageEnglish (US)
Article numbere0115639
JournalPloS one
Volume10
Issue number1
DOIs
StatePublished - Jan 28 2015
Externally publishedYes

ASJC Scopus subject areas

  • General

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