Directed actin assembly and motility

Methods Enzymol. 2014:540:283-300. doi: 10.1016/B978-0-12-397924-7.00016-9.

Abstract

The actin cytoskeleton is a key component of the cellular architecture. However, understanding actin organization and dynamics in vivo is a complex challenge. Reconstitution of actin structures in vitro, in simplified media, allows one to pinpoint the cellular biochemical components and their molecular interactions underlying the architecture and dynamics of the actin network. Previously, little was known about the extent to which geometrical constraints influence the dynamic ultrastructure of these networks. Therefore, in order to study the balance between biochemical and geometrical control of complex actin organization, we used the innovative methodologies of UV and laser patterning to design a wide repertoire of nucleation geometries from which we assembled branched actin networks. Using these methods, we were able to reconstitute complex actin network organizations, closely related to cellular architecture, to precisely direct and control their 3D connections. This methodology mimics the actin networks encountered in cells and can serve in the fabrication of innovative bioinspired systems.

Keywords: Actin; Actin-based 3D connections; Arp2/3 complex; Micropatterning; Motility; Self-organized networks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / metabolism*
  • Actin Cytoskeleton / ultrastructure*
  • Actins / chemistry
  • Actins / metabolism*
  • Actins / ultrastructure*
  • Animals
  • Equipment Design
  • Lasers
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Polymerization
  • Ultraviolet Rays

Substances

  • Actins