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Focal Adhesion Induction at the Tip of a Functionalized Nanoelectrode

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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.

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Authors and Affiliations

  1. Department of Bioengineering, The Pennsylvania State University, 205 Hallowell Building, University Park, PA, 16802, USA

    Daniela E. Fuentes & Peter J. Butler

  2. Center for Single Molecule Biophysics, University at Buffalo, State University of New York, 301 Cary Hall, Buffalo, NY, 14214, USA

    Chilman Bae

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  1. Daniela E. Fuentes
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  2. Chilman Bae
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Correspondence to Peter J. Butler.

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Associate Editor Edward Guo oversaw the review of this article.

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Fuentes, D.E., Bae, C. & Butler, P.J. Focal Adhesion Induction at the Tip of a Functionalized Nanoelectrode. Cel. Mol. Bioeng. 4, 616–626 (2011). https://doi.org/10.1007/s12195-011-0214-7

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