How to cite this article: Meseke M, Förster E. 2013. A 3D-matrigel/microbead assay for the visualization of mechanical tractive forces at the neurite-substrate interface of cultured neurons. J Biomed Mater Res Part A 2013:101A:1726–1733.
A 3D-matrigel/microbead assay for the visualization of mechanical tractive forces at the neurite–substrate interface of cultured neurons †
Article first published online: 27 NOV 2012
Copyright © 2012 Wiley Periodicals, Inc.
Journal of Biomedical Materials Research Part A
Volume 101A, Issue 6, pages 1726–1733, June 2013
How to Cite
Meseke, M. and Förster, E. (2013), A 3D-matrigel/microbead assay for the visualization of mechanical tractive forces at the neurite–substrate interface of cultured neurons . J. Biomed. Mater. Res., 101A: 1726–1733. doi: 10.1002/jbm.a.34477
- Issue published online: 13 APR 2013
- Article first published online: 27 NOV 2012
- Manuscript Accepted: 6 OCT 2012
- Manuscript Revised: 24 SEP 2012
- Manuscript Received: 8 JUL 2012
- Deutsche Forschungsgemeinschaft (FO 223/6-1 to EF)
- neuronal migration;
- soma translocation;
Mechanical properties of neuronal processes contribute to neuronal function, to resistance of fiber tracts against mechanical trauma, and to morphological changes during development and neurodegeneration. Conventional in vitro cell culture systems on inflexible substrates do not allow for the visualization of changing mechanical stress between neurites and their substrate. To solve this problem, we adapted a three-dimensional gel matrix assay to visualize mechanical traction forces at the neurite–substrate interface. We chose matrigel as substrate because in this matrix various types of neurons initially adapt a bipolar morphology while migrating, similar to migrating neurons in vivo. To visualize emerging traction forces between neurites and their substrate, microbeads were embedded into the matrix as visible landmarks. We first analyzed mechanical distortion of matrigel by stepwise movements of a glass pipette tip under control of a micromanipulator to ensure reproducibility of induced bead displacement. The assay was then used to study the effect of the microtubule disrupting drug nocodazole on neuronal processes. By monitoring displacement of matrigel-embedded microbeads, we visualized here for the first time emerging mechanical traction forces between the leading process and the substrate during nocodazole-induced soma translocation. We did not observe bead displacement by processes of aged neurons. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.