Substrate topography determines neuronal polarization and growth in vitro

PLoS One. 2013 Jun 13;8(6):e66170. doi: 10.1371/journal.pone.0066170. Print 2013.

Abstract

The establishment of neuronal connectivity depends on the correct initial polarization of the young neurons. In vivo, developing neurons sense a multitude of inputs and a great number of molecules are described that affect their outgrowth. In vitro, many studies have shown the possibility to influence neuronal morphology and growth by biophysical, i.e. topographic, signaling. In this work we have taken this approach one step further and investigated the impact of substrate topography in the very early differentiation stages of developing neurons, i.e. when the cell is still at the round stage and when the first neurite is forming. For this purpose we fabricated micron sized pillar structures with highly reproducible feature sizes, and analyzed neurons on the interface of flat and topographic surfaces. We found that topographic signaling was able to attract the polarization markers of mouse embryonic neurons -N-cadherin, Golgi-centrosome complex and the first bud were oriented towards topographic stimuli. Consecutively, the axon was also preferentially extending along the pillars. These events seemed to occur regardless of pillar dimensions in the range we examined. However, we found differences in neurite length that depended on pillar dimensions. This study is one of the first to describe in detail the very early response of hippocampal neurons to topographic stimuli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism
  • Cadherins / metabolism
  • Cell Differentiation
  • Cell Polarity*
  • Cells, Cultured
  • Centrosome / metabolism
  • Golgi Apparatus / metabolism
  • Growth Cones / metabolism
  • Mice
  • Neurites / metabolism
  • Neurons / cytology*
  • Neurons / physiology*
  • Phosphorylation
  • Protein Transport
  • Rats
  • Signal Transduction
  • Tyrosine / metabolism

Substances

  • Cadherins
  • Tyrosine

Grants and funding

Part of this work was supported by the Flanders Fund for Scientific Research (FWO G 0.666.10N, www.fwo.be), the Federal Office for Scientific Affairs (IUAP p6/43), Flemish Government Methusalem Grant and Spanish Ministry of Science and Innovation Ingenio-Consolider CSD2010-00064 and SAF2010-14906 (CGD). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.