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A versatile prion replication assay in organotypic brain slices.

Jeppe Falsig1, Christian Julius, Ilan Margalith

  • 1Institute of Neuropathology, University of Zürich, Schmelzbergstrasse 12, Zürich, Switzerland.

Nature Neuroscience
|December 11, 2007
PubMed
Summary

A new prion organotypic slice culture assay (POSCA) amplifies prion protein (PrPSc) ex vivo, closely mimicking in vivo infection. This method offers a faster, effective tool for studying prion diseases and their potential treatments.

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Area of Science:

  • Neuroscience
  • Infectious Diseases
  • Biotechnology

Background:

  • Prion diseases are fatal neurodegenerative disorders.
  • Studying prions ex vivo is crucial but challenging due to limited methods.
  • Existing methods often fail to support replication of diverse prion strains.

Purpose of the Study:

  • To develop a novel ex vivo method for prion replication and titration.
  • To establish a model that closely mimics intracerebral prion infection.
  • To investigate the role of microglia in prion disease progression.

Main Methods:

  • Development of a prion organotypic slice culture assay (POSCA).
  • Infection of mouse cerebellar slices with prions.
  • Quantification of abnormal prion protein (PrPSc) accumulation.

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  • Pharmacogenetic ablation of microglia to assess their role.

    • Main Results:

    • POSCA achieved >10(5)-fold PrPSc amplification within 35 days, comparable to in vivo levels but fivefold faster.
    • PrPSc accumulation occurred predominantly in the molecular layer, mirroring in vivo infection.
    • The assay detected prion strains from bovine and ovine sources with variable efficiency.
    • Microglia ablation increased prion titers by 15-fold, suggesting a defensive role for microglia.

    Conclusions:

    • POSCA is an effective ex vivo tool for prion amplification and titration.
    • Microglial activation in prion diseases may represent an innate defense mechanism.
    • This assay provides a valuable platform for studying prion pathogenesis and therapeutic interventions.