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Related Experiment Videos

Neuronal apoptosis studied by a sequential TUNEL technique: a method for tract-tracing.

A Hara1, M Niwa, T Iwai

  • 1Department of Pathology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu 500-8705, Japan. ahara@cc.gifu-u.ac.jp

Brain Research. Brain Research Protocols
|August 14, 1999
PubMed
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This study introduces a novel tract-tracing method using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) to track apoptotic neurons. The technique traces fragmented DNA in neuronal fibers, revealing neuroanatomical connections in gerbil hippocampal CA1 neurons.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Neuroanatomy

Background:

  • Traditional neuroanatomical studies rely on exogenous tracers (xenobiotics).
  • A method to trace neuronal connections using intrinsic cellular markers is needed.
  • Apoptotic processes involve DNA fragmentation and can utilize axonal transport mechanisms.

Purpose of the Study:

  • To develop and describe a novel tract-tracing technique using an intrinsic marker.
  • To identify and trace neuronal projections from apoptotic cells.
  • To investigate neuroanatomical connections in the gerbil hippocampus following ischemia.

Main Methods:

  • Sequential in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) of DNA fragments.
  • Detection of fragmented DNA transported within neuronal fibers.

Related Experiment Videos

  • Confirmation of DNA transport using DAPI fluorescence staining.
  • Time-course analysis of TUNEL-positive DNA movement.

    • Main Results:

    • The TUNEL method successfully identified fragmented DNA in neuronal fibers of gerbil hippocampal CA1 neurons after transient forebrain ischemia.
    • Transported DNA was confirmed using DAPI staining.
    • The technique traced apoptotic neurons to the ends of their neuronal fibers, demonstrating projections.

    Conclusions:

    • The described TUNEL-based method offers a novel approach for tract-tracing using an intrinsic marker of apoptosis.
    • This technique allows for the visualization of neuronal projections from selected apoptotic cells.
    • The method highlights the utility of studying apoptotic processes for understanding neuroanatomy.