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Related Experiment Videos

Dynamics of the neuronal intermediate filaments

S Okabe1, H Miyasaka, N Hirokawa

  • 1Department of Anatomy and Cell Biology, School of Medicine, University of Tokyo, Japan.

The Journal of Cell Biology
|April 1, 1993
PubMed
Summary

Neuronal intermediate filaments in growing axons exhibit slow turnover, with new subunits incorporated laterally and segmentally. This process is regulated by axonal growth, as shown by fluorescence recovery studies.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Neuronal intermediate filaments, such as neurofilaments, are crucial for maintaining neuronal structure and function.
  • Understanding the dynamics and turnover of these filaments is essential for comprehending axonal development and maintenance.

Purpose of the Study:

  • To investigate the dynamics and turnover mechanisms of neuronal intermediate filaments in living neurons.
  • To determine if optical methods used for analysis perturb axonal growth or filament dynamics.

Main Methods:

  • Utilized photobleaching of fluorescently-labeled neurofilament L protein in mouse sensory neuron axons.
  • Employed immunoelectron microscopy to identify incorporation sites of biotinylated neurofilament L protein.
  • Performed low-light-level imaging to minimize potential perturbations.

Main Results:

  • Photobleaching and imaging did not significantly affect axonal growth rate or filament addition.
  • Slow fluorescence recovery (half-time of 40 min) was observed post-photobleaching.
  • Faster recovery in growing axons suggests growth-dependent turnover regulation.
  • Biotinylated neurofilament L incorporation occurred at discrete sites, gradually forming continuous arrays.

Conclusions:

  • Neuronal intermediate filaments in growing axons undergo turnover within the axoplasm.
  • A mechanism involving lateral and segmental incorporation of new subunits likely drives this turnover.
  • Axonal growth actively regulates the rate of neurofilament dynamics.

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