A morphometric screen identifies specific roles for microtubule-regulating genes in neuronal development of P19 stem cells

PLoS One. 2013 Nov 18;8(11):e79796. doi: 10.1371/journal.pone.0079796. eCollection 2013.

Abstract

The first morphological change after neuronal differentiation is the microtubule-dependent initiation of thin cell protrusions called neurites. Here we performed a siRNA-based morphometric screen in P19 stem cells to evaluate the role of 408 microtubule-regulating genes during this early neuromorphogenesis step. This screen uncovered several novel regulatory factors, including specific complex subunits of the microtubule motor dynein involved in neurite initiation and a novel role for the microtubule end-binding protein EB2 in attenuation of neurite outgrowth. Epistasis analysis suggests that competition between EB1 and EB2 regulates neurite length, which links its expression to neurite outgrowth. We propose a model that explains how microtubule regulators can mediate cellular morphogenesis during the early steps of neuronal development by controlling microtubule stabilization and organizing dynein-generated forces.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cytoplasm / genetics
  • Cytoplasm / metabolism
  • Dynactin Complex
  • Dyneins / genetics
  • Dyneins / metabolism
  • Mice
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / genetics*
  • Microtubules / metabolism*
  • Neurites / metabolism
  • Neurogenesis / genetics*
  • Neurons / cytology*
  • Neurons / metabolism
  • Protein Binding / genetics
  • Stem Cells / cytology*
  • Stem Cells / metabolism*

Substances

  • Dynactin Complex
  • Microtubule-Associated Proteins
  • Dyneins

Grants and funding

This work was funded by the FORSYS partner initiative of the German Federal Ministry of Education and Research (BMBF, grant 0315258) to LD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.