Focal adhesion induction at the tip of a functionalized nanoelectrode

Daniela E. Fuentes, Chilman Bae, Peter J. Butler

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Cells dynamically interact with their physical micro-environment through the assembly of nascent focal contacts and focal adhesions. The dynamics and mechanics of these contact points are controlled by transmembrane integrins and an array of intracellular adaptor proteins. In order to study the mechanics and dynamics of focal adhesion assembly, we have developed a technique for the timed induction of a nascent focal adhesion. Bovine aortic endothelial cells were approached at the apical surface by a nanoelectrode whose position was controlled with a resolution of 10 s of nanometers using changes in electrode current to monitor distance from the cell surface. Since this probe was functionalized with fibronectin, a focal contact formed at the contact location. Nascent focal adhesion assembly was confirmed using time-lapse confocal fluorescent images of red fluorescent protein-tagged talin, an adapter protein that binds to activated integrins. Binding to the cell was verified by noting a lack of change of electrode current upon retraction of the electrode. This study demonstrates that functionalized nanoelectrodes can enable precisely-timed induction and 3-D mechanical manipulation of focal adhesions and the assay of the detailed molecular kinetics of their assembly.

Original languageEnglish (US)
Pages (from-to)616-626
Number of pages11
JournalCellular and Molecular Bioengineering
Volume4
Issue number4
DOIs
StatePublished - Dec 2011

All Science Journal Classification (ASJC) codes

  • Modeling and Simulation
  • Biochemistry, Genetics and Molecular Biology(all)

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