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Microtubule plus-end conformations and dynamics in the periphery of interphase mouse fibroblasts

Sandra Zovko1, Jan Pieter Abrahams, Abraham J Koster

  • 1Section Electron Microscopy, Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands. zovko.sandra@gmail.com

Molecular Biology of the Cell
|May 16, 2008
PubMed

Insights

Microtubule (MT) plus-end dynamics were studied in mouse fibroblasts. Nine distinct conformations were identified, with their frequency influenced by cell periphery proximity, suggesting a link between structure and dynamic instability.

Area of Science:

  • Cell Biology
  • Cytoskeleton Dynamics
  • Microtubule Structure

Background:

  • Microtubules (MTs) exhibit dynamic instability, alternating between growth, pause, and shrinkage.
  • In vitro studies link MT plus-end conformation to dynamic states (frayed, blunt, sheet).
  • In situ conformations and their dynamic states require further investigation.

Purpose of the Study:

  • Investigate the dynamics of MT plus ends in mouse fibroblasts.
  • Identify and characterize distinct MT plus-end conformations in situ.
  • Determine the relationship between MT plus-end conformation and dynamic state.

Main Methods:

  • Cryo-fixation, freeze-substitution, and flat-embedding of mouse fibroblasts (3T3s).
  • Electron microscopy and tomography for high-resolution imaging of MT plus ends.
  • Nocodazole treatment to induce MT depolymerization and analyze dynamic transitions.

Main Results:

  • Identified nine morphologically distinct MT plus-end conformations in vivo.
  • Conformation frequency correlated with proximity to the cell border, indicating peripheral influence.
  • Established a transition pathway from growth to shrinkage via sheet-frayed and flared ends.
  • Developed a kinetic model for MT depolymerization dynamics.

Conclusions:

  • MT plus-end conformation is linked to its dynamic state in vivo.
  • Cell periphery features influence MT dynamic instability.
  • A novel transition pathway and kinetic model for MT depolymerization were proposed.

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