title: Two distinct filopodia populations at the growth cone allow to sense nanotopographical extracellular matrix cues to guide neurite outgrowth.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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 behavior 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.
  
  
  CONCLUSIONS
  We propose that a constant crosstalk between both filopodia populations allows stochastic sensing of nanotopographical 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.
authors.affiliation: Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul, Republic of Korea.
authors.forename: Kyung-Jin
authors.initials: KJ
authors.lastname: Jang
authors.forename: Min Sung
authors.initials: MS
authors.lastname: Kim
authors.forename: Daniel
authors.initials: D
authors.lastname: Feltrin
authors.forename: Noo Li
authors.initials: NL
authors.lastname: Jeon
authors.forename: Kahp-Yang
authors.initials: KY
authors.lastname: Suh
authors.forename: Olivier
authors.initials: O
authors.lastname: Pertz
mesh_terms: D001921:Brain / Q000502:physiology; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism*; D020439:Growth Cones / Q000378:metabolism*; D006801:Humans; D007211:Indoles / Q000737:chemistry; D007797:Laminin / Q000737:chemistry* / Q000378:metabolism; D008858:Microscopy, Phase-Contrast / Q000379:methods; D016501:Neurites / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D011554:Pseudopodia / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry
chemical_list: D007211:Indoles; D007797:Laminin; D014404:Tubulin; C007293:DAPI