Autophagy and cell death of Purkinje cells overexpressing Doppel in Ngsk Prnp-deficient mice

Brain Pathol. 2010 Jan;20(1):119-32. doi: 10.1111/j.1750-3639.2008.00245.x. Epub 2008 Nov 24.

Abstract

In Ngsk prion protein (PrP)-deficient mice (NP(0/0)), ectopic expression of PrP-like protein Doppel (Dpl) in central neurons induces significant Purkinje cell (PC) death resulting in late-onset ataxia. NP(0/0) PC death is partly prevented by either knocking-out the apoptotic factor BAX or overexpressing the anti-apoptotic factor BCL-2 suggesting that apoptosis is involved in Dpl-induced death. In this study, Western blotting and immunohistofluorescence show that both before and during significant PC loss, the scrapie-responsive gene 1 (Scrg1)--potentially associated with autophagy--and the autophagic markers LC3B and p62 increased in the NP(0/0) PCs whereas RT-PCR shows stable mRNA expression, suggesting that the degradation of autophagic products is impaired in NP(0/0) PCs. At the ultrastructural level, autophagic-like profiles accumulated in somatodendritic and axonal compartments of NP(0/0), but not wild-type PCs. The most robust autophagy was observed in NP(0/0) PC axon compartments in the deep cerebellar nuclei suggesting that it is initiated in these axons. Our previous and present data indicate that Dpl triggers autophagy and apoptosis in NP(0/0) PCs. As observed in amyloid neurodegenerative diseases, upregulation of autophagic markers as well as extensive accumulation of autophagosomes in NP(0/0) PCs are likely to reflect a progressive dysfunction of autophagy that could trigger apoptotic cascades.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy
  • Axons / pathology
  • Axons / ultrastructure
  • Blotting, Western
  • Cell Death
  • Cerebellar Cortex / pathology
  • Cerebellar Cortex / ultrastructure
  • Cerebellar Nuclei / pathology
  • Cerebellar Nuclei / ultrastructure
  • Cerebellum / cytology
  • Cerebellum / drug effects
  • Cerebellum / metabolism
  • Dendrites / pathology
  • Dendrites / ultrastructure
  • Fluorescent Antibody Technique
  • GPI-Linked Proteins
  • Genotype
  • Immunohistochemistry
  • Lysosomal Membrane Proteins / biosynthesis
  • Lysosomal Membrane Proteins / genetics
  • Mice
  • Mice, Knockout
  • Microtubule-Associated Proteins / biosynthesis
  • Microtubule-Associated Proteins / genetics
  • Nerve Tissue Proteins / biosynthesis
  • Nerve Tissue Proteins / genetics
  • Prions / biosynthesis
  • Prions / genetics*
  • Purkinje Cells / metabolism*
  • Purkinje Cells / pathology*
  • Purkinje Cells / ultrastructure
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor TFIIH
  • Transcription Factors / biosynthesis
  • Transcription Factors / genetics
  • gamma-Aminobutyric Acid / metabolism

Substances

  • GPI-Linked Proteins
  • Gtf2h1 protein, mouse
  • Lamp1 protein, mouse
  • Lysosomal Membrane Proteins
  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Nerve Tissue Proteins
  • Prions
  • Prnd protein, mouse
  • Scrg1 protein, mouse
  • Transcription Factors
  • Transcription Factor TFIIH
  • gamma-Aminobutyric Acid