Authors

Jang, K.-J. J., Kim, M. S., Feltrin, D., Jeon, N. L., Suh, K.-Y. Y., & Pertz, O.

Affiliations

Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul, Republic of Korea, School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, Republic of Korea, Department of Biomedicine, Institute for Biochemistry and Genetics, University of Basel, Basel, Switzerland.

Application Area

Neuroscience

Time-Lapse Microscopy

Abstract

The process of neurite outgrowth is the initial step in producing the neuronal processes that wire the brain. Current models about neurite outgrowth have been derived from classic two-dimensional (2D) cell culture systems, which do not recapitulate the topographical cues that are present in the extracellular matrix (ECM) in vivo. Here, we explore how ECM nanotopography influences neurite outgrowth. We show that, when the ECM protein laminin is presented on a line pattern with nanometric size features, it leads to orientation of neurite outgrowth along the line pattern. This is also coupled with a robust increase in neurite length. The sensing mechanism that allows neurite orientation occurs through a highly stereotypical growth cone behaviour involving two filopodia populations. Non-aligned filopodia on the distal part of the growth cone scan the pattern in a lateral back and forth motion and are highly unstable. Filopodia at the growth cone tip. Align with the line substrate, are stabilized by an F-actin rich cytoskeleton and enable steady neurite extension. This stabilization event most likely occurs by integration of signals emanating from non-aligned and aligned filopodia which sense different extent of adhesion surface on the line pattern. In contrast, on the 2D substrate only unstable filopodia are observed at the growth cone, leading to frequent neurite collapse events and less efficient outgrowth. We propose that a constant crosstalk between both filopodia populations allows stochastic sensing of anotopographical ECM cues, leading to oriented and steady neurite outgrowth. Our work provides insight in how neuronal growth cones can sense geometric ECM cues. This has not been accessible previously using routine 2D culture systems.

Extract

… “steered by Metamorph software (Molecular Devices) and using a LED-based light source (Coolled)”…

Product Associated Features

pE-100: A range of compact, simple-to-use, single wavelength illumination systems for screening fluorescence.

Diascopic Technique

Phase Contrast

Live Cell Issues

-

Product Type

pE-100

Journal

PloS one

Year of Publication

2010

Country of Publication

South Korea