A versatile prion replication assay in organotypic brain slices

Falsig, Jeppe; Julius, Christian; Margalith, Ilan; Schwarz, Petra; Heppner, Frank L.; Aguzzi, Adriano

doi:10.1038/nn2028

Cited by 131 publications

(161 citation statements)

References 32 publications

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“…The absence of effect of TREM2 on Aβ plaques and prion load is surprising; it suggests that the microglial activation state may not necessarily correlate with their phagocytic capacity. Together with our previous findings that depletion of microglia enhances prion replication in organotypic cerebellar slices (Falsig, et al, 2008), we hypothesize that the availability of microglia, and perhaps a basal level of activation, may be crucial for phagocytosis, whereas further activation does not appear to augment the clearance of prions. Accordingly, lipopolysaccharide-stimulated microglial activation did not enhance further phagocytosis of prions (Hughes, et al, 2010).…”

Section: Discussionsupporting

confidence: 80%

“…Moreover, PrP Sc deposition and astrogliosis were similar in all three groups. Since the phagocytic activity of microglia is a well-established limiting factor for prion propagation within the brain (Falsig, et al, 2008,Kranich, et al, 2010, the unchanged PrP Sc deposition and prion infectivity titer suggested that the phagocytosis of prions by TREM2 -/-microglia was not markedly affected. However, TREM2 seems to be involved in microglial activation, as the prion-induced enhancement of IBA-1 reactivity was attenuated in TREM2 -/-mice.…”

Section: Discussionmentioning

confidence: 99%

“…Cerebellar organotypic slices were prepared from 9-11-day old pups and maintained according to previously published protocols (Falsig, et al, 2008). In Tg20/CD11b-HSVTK brain slices, microglia were depleted by adding ganciclovir from 0 to 21 days in vitro.…”

Section: Organotypic Slice Cultures and Microglia Depletionmentioning

confidence: 99%

“…It was then found that microglia, the most prevalent immune cells of the brain, act as the main scavenger of prions. In prion-infected cerebellar organotypic cultured slices, depletion of microglia vastly enhances prion deposition and leads to increased prion infectivity (Falsig, et al, 2008). The opsonin Mfge8 is instrumental for microglia-dependent clearance of prions, possibly by bridging phospholipids trapped within PrP Sc to integrins expressed by microglia (Kranich, et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Triggering receptor expressed on myeloid cells-2 is involved in prion-induced microglial activation but does not contribute to prion pathogenesis in mouse brains

Zhu

Herrmann

et al. 2015

Neurobiology of Aging

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Dysfunctional variants of the innate immune cell surface receptor TREM2 (triggering receptor expressed on myeloid cells-2) were identified as major genetic risk factors for Alzheimer's disease and other neurodegenerative conditions. Here we assessed a possible involvement of TREM2 in prion disease. We report that TREM2 expression by microglia is significantly up-regulated upon prion infection. However, depletion of TREM2 did not affect disease incubation time and survival after intracerebral prion infection. Interestingly, markers of microglial activation were attenuated in prion-infected TREM2(-/-) mice, suggesting an involvement of TREM2 in prion-induced microglial activation. Further phenotype profiling of microglia revealed that TREM2 deficiency did not change microglial phenotypes. We conclude that TREM2 is involved in prion-induced microglial activation but does not noticeably modulate the pathogenesis of experimental prion infections. | P a g e AbstractDysfunctional variants of the innate immune cell surface receptor TREM2 (triggering receptor expressed on myeloid cells-2) were identified as major genetic risk factors for Alzheimer's disease and other neurodegenerative conditions. Here we assessed a possible involvement of TREM2 in prion disease. We report that TREM2 expression by microglia is significantly upregulated upon prion infection. However, depletion of TREM2 did not affect disease incubation time and survival after intracerebral prion infection. Interestingly, markers of microglial activation were attenuated in prion-infected TREM2 -/-mice, suggesting an involvement of TREM2 in prion-induced microglial activation. Further phenotype profiling of microglia revealed that TREM2 deficiency did not change microglial phenotypes. We conclude that TREM2 is involved in prion-induced microglial activation, but does not noticeably modulate the pathogenesis of experimental prion infections.

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Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Organotypic Slice Cultures and Microglia Depletionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Triggering receptor expressed on myeloid cells-2 is involved in prion-induced microglial activation but does not contribute to prion pathogenesis in mouse brains

Zhu

Herrmann

et al. 2015

Neurobiology of Aging

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“…For microglia depletion experiments, tga20 TK+ slices were treated with ganciclovir (GCV, 5 μg ml -1 ) for 14 days prior to antibody treatment. At this time point less than 1% of microglia were left in the tissue 39 . …”

Section: Manganese-enhanced Magnetic Resonance Imaging (Memri)mentioning

confidence: 90%

The toxicity of antiprion antibodies is mediated by the flexible tail of the prion protein

Sonati

Reimann

Falsig

et al. 2013

Nature

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Prion infections cause lethal neurodegeneration. This process requires the cellular prion protein (PrP(C); ref. 1), which contains a globular domain hinged to a long amino-proximal flexible tail. Here we describe rapid neurotoxicity in mice and cerebellar organotypic cultured slices exposed to ligands targeting the 1 and 3 helices of the PrP(C) globular domain. Ligands included seven distinct monoclonal antibodies, monovalent Fab1 fragments and recombinant single-chain variable fragment miniantibodies. Similar to prion infections, the toxicity of globular domain ligands required neuronal PrP(C), was exacerbated by PrP(C) overexpression, was associated with calpain activation and was antagonized by calpain inhibitors. Neurodegeneration was accompanied by a burst of reactive oxygen species, and was suppressed by antioxidants. Furthermore, genetic ablation of the superoxide-producing enzyme NOX2 (also known as CYBB) protected mice from globular domain ligand toxicity. We also found that neurotoxicity was prevented by deletions of the octapeptide repeats within the flexible tail. These deletions did not appreciably compromise globular domain antibody binding, suggesting that the flexible tail is required to transmit toxic signals that originate from the globular domain and trigger oxidative stress and calpain activation. Supporting this view, various octapeptide ligands were not only innocuous to both cerebellar organotypic cultured slices and mice, but also prevented the toxicity of globular domain ligands while not interfering with their binding. We conclude that PrP(C) consists of two functionally distinct modules, with the globular domain and the flexible tail exerting regulatory and executive functions, respectively. Octapeptide ligands also prolonged the life of mice expressing the toxic PrP(C) mutant, PrP(Δ94-134), indicating that the flexible tail mediates toxicity in two distinct PrP(C)-related conditions. Flexible tail-mediated toxicity may conceivably play a role in further prion pathologies, such as familial Creutzfeldt-Jakob disease in humans bearing supernumerary octapeptides. (Fig. 1b). None of three high-affinity antibodies to the octapeptide repeats (OR, residues 50-90 embedded within the FT) were neurotoxic (Fig. 1b). Antibodies POM3 and D13, which bind the "charged cluster-2" 11 (CC2, residues 95-110), were innocuous at 67 nM but neurotoxic at 200 nM (Fig. 1b). None of the tested antibodies were toxic to Prnp o/o COCS ( Supplementary Fig. 2a). The identity of the targeted epitopes appeared to be a better predictor of PrP C antibody toxicity than their affinity to PrP C , suggesting that neurotoxicity resulted from the interaction of antibodies with specific PrP C domains (Supplementary Table 2).The mechanisms of neurotoxicity were further explored using POM1, a highly toxic antibody targeting the GD. Wild-type (wt) and tga20 COCS lost most granule cells (CGC) within 28 and 14 days post-exposure (dpe) to POM1, respectively (Fig. 2a-c). Controls included POM1-treated Prnp o/o COCS 12 , t...

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Prions

Lawson¹

2015

Encyclopedia of Life Sciences

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Prion diseases are invariably fatal neurodegenerative disorders associated with the aberrant folding of the normal cellular prion protein. The disease affects both humans and animals and in humans occurs in sporadic, familial and acquired forms. In the absence of a conventional infectious agent, the acquired forms of the disease occur through the transmission of the misfolded form of the prion protein, or prion. This article highlights the evidence for the principle of a ‘protein‐only’ transmissible disease and addresses some of the controversies including the existence of prion strains and the species barrier. It will also consider the expanding spectrum of ‘prion‐like’ diseases. Key Concepts An overview of prion diseases that affect humans and animals. Biology of the cellular prion protein. Prion protein misfolding and disease. Proving the ‘protein‐only’ hypothesis. The molecular basis of the species barrier and prion strains. An introduction to ‘prion‐like’ protein aggregation.

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A versatile prion replication assay in organotypic brain slices

Cited by 131 publications

References 32 publications

Triggering receptor expressed on myeloid cells-2 is involved in prion-induced microglial activation but does not contribute to prion pathogenesis in mouse brains

Triggering receptor expressed on myeloid cells-2 is involved in prion-induced microglial activation but does not contribute to prion pathogenesis in mouse brains

The toxicity of antiprion antibodies is mediated by the flexible tail of the prion protein

Prions

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