Strain‐Associated Variations in Abnormal PrP Trafficking of Sheep Scrapie

Jeffrey, Martin; McGovern, Gillian; Goodsir, C. M.; González, Lorenzo

doi:10.1111/j.1750-3639.2008.00150.x

Cited by 35 publications

(56 citation statements)

References 47 publications

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“…In addition to labeling for PrP, immunogold labeling for ubiquitin was also performed, as previously described. 9 Sections were viewed in a Jeol 1200 EX electron microscope.…”

Section: Electron Microscopic Immunohistochemical Proceduresmentioning

confidence: 99%

“…8 -10 Such PrP d accumulations specifically co-localize with several kinds of cellular pathology, including abnormal endocytic structures, microfolding of the plasma membrane, and excess lysosomes. 9 In several murine scrapie models (eg, ME7), there is also conspicuous synaptic loss (not co-localized with PrP d ), as well as marked degeneration of axons. 11 Tg(PG14) mice express the murine homologue of PrP carrying a nine-octapeptide insertional mutation described in human patients with an inherited prion disease.…”

mentioning

confidence: 99%

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Prion Protein with an Insertional Mutation Accumulates on Axonal and Dendritic Plasmalemma and Is Associated with Distinctive Ultrastructural Changes

Jeffrey¹,

Goodsir²,

McGovern³

et al. 2009

The American Journal of Pathology

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Prion diseases are fatal neurological diseases characterized by central nervous system deposition of abnormal forms of a membrane glycoprotein designated PrP (prion protein). Tg(PG14) transgenic mice express PrP that harbor a nine-octapeptide insertional mutation homologous to one described in a familial prion disease of humans. Tg(PG14) mice spontaneously develop a fatal neurological illness accompanied by massive apoptosis of cerebellar granule neurons and accumulation of an aggregated and weakly protease-resistant form of PrP that is not infectious. Previous light microscopic analyses of these mice left open questions regarding the subcellular distribution of the mutant protein and the nature of the neuropathological lesions produced. To address these questions, we undertook an immunogold electron microscopic study of Tg(PG14) mice. We found that mutant PrP is localized primarily on the plasma membrane of dendrites and unmyelinated axons in the hippocampus and cerebellum, with little labeling of either neuronal cell bodies or intracellular organelles. PrP deposits were shown to be associated with degenerative changes in dendritic structure. We also describe for the first time marked pathology in myelinated axons , and alterations in the axon/oligodendrocyte interface. Taken together , our results suggest cellular mechanisms by which mutant PrPs produce pathology. In addition, they highlight distinctions between familial and infectious prion disorders at the ultrastructural level that correlate with differences in cellular trafficking of the disease-

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“…In addition to labeling for PrP, immunogold labeling for ubiquitin was also performed, as previously described. 9 Sections were viewed in a Jeol 1200 EX electron microscope.…”

Section: Electron Microscopic Immunohistochemical Proceduresmentioning

confidence: 99%

mentioning

confidence: 99%

Prion Protein with an Insertional Mutation Accumulates on Axonal and Dendritic Plasmalemma and Is Associated with Distinctive Ultrastructural Changes

Jeffrey¹,

Goodsir²,

McGovern³

et al. 2009

The American Journal of Pathology

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“…Distinguishing between these different possibilities will require further investigation of Sup35-GFP GPI aggregates. TSE-associated GPI-anchored aggregates composed of misfolded prion protein usually appear as small, diffuse assemblies that do not appear to be amyloid fibrils (14,15,22). However, PrPSc can also form amyloid, demonstrated by the presence of extracellular plaques in some cases of TSE and in mice genetically modified to express anchorless prion protein, following extraction or release from cell membranes and in vitro (26,37,(82)(83)(84).…”

Section: Discussionmentioning

confidence: 99%

“…The cross-␤ arrangement yields characteristic reflections by fiber diffraction and typically stains with the amyloid-specific dyes Thioflavin T or S and Congo Red (17)(18)(19)(20). In most instances, PrPSc is membrane-bound as granular, diffuse, nonfibrillar structures, with amyloid plaques only manifesting in certain TSE diseases, such as kuru, Gerstmann-StrausslerScheinker disease, variant Creutzfeldt-Jakob disease, and a minority of sporadic Creutzfeldt-Jakob disease cases (14,(21)(22)(23)(24)(25). The effect of GPI anchoring of PrP on TSE pathogenesis and infectivity has been investigated using a transgenic mouse that expresses PrPC lacking a GPI anchor (26,27).…”

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confidence: 99%

Glycosylphosphatidylinositol Anchoring Directs the Assembly of Sup35NM Protein into Non-fibrillar, Membrane-bound Aggregates

Marshall¹,

Offerdahl²,

Speare

et al. 2014

Journal of Biological Chemistry

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Background: Sup35NM is a soluble protein that when misfolded forms amyloid fibril aggregates. Results: When tethered to membranes via a lipid anchor, Sup35NM aggregates adopt a non-fibrillar, membrane-bound structure. Conclusion: Lipid anchor-directed membrane association prevents assembly into fibrils. Significance: This may explain differences between prion diseases compared with related protein aggregation diseases because only prion diseases involve aggregation of a lipid-anchored protein.

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“…The PrP res aggregates in the two inocula have different ultrastructures, with fibrils adopting an amyloid conformation and microsomebound aggregates adopting a nonfibrillar, apparently nonamyloid structure (7,35,44,49,(118)(119)(120)(121). Furthermore, PrP res in the two inocula might be expected to interact differently with distinct membrane regions.…”

Section: Discussionmentioning

confidence: 99%

PrP Knockout Cells Expressing Transmembrane PrP Resist Prion Infection

Marshall

Hughson

Vascellari

et al. 2017

J Virol

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Glycosylphosphatidylinositol (GPI) anchoring of the prion protein (PrP C ) influences PrP C misfolding into the disease-associated isoform, PrP res , as well as prion propagation and infectivity. GPI proteins are found in cholesterol-and sphingolipid-rich membrane regions called rafts. Exchanging the GPI anchor for a nonraft transmembrane sequence redirects PrP C away from rafts. Previous studies showed that nonraft transmembrane PrP C variants resist conversion to PrP res when transfected into scrapie-infected N2a neuroblastoma cells, likely due to segregation of transmembrane PrP C and GPI-anchored PrP res in distinct membrane environments. Thus, it remained unclear whether transmembrane PrP C might convert to PrP res if seeded by an exogenous source of PrP res not associated with host cell rafts and without the potential influence of endogenous expression of GPI-anchored PrP C . To further explore these questions, constructs containing either a C-terminal wild-type GPI anchor signal sequence or a nonraft transmembrane sequence containing a flexible linker were expressed in a cell line derived from PrP knockout hippocampal neurons, NpL2. NpL2 cells have physiological similarities to primary neurons, representing a novel and advantageous model for studying transmissible spongiform encephalopathy (TSE) infection. Cells were infected with inocula from multiple prion strains and in different biochemical states (i.e., membrane bound as in brain microsomes from wild-type mice or purified GPI-anchorless amyloid fibrils). Only GPI-anchored PrP C supported persistent PrP res propagation. Our data provide strong evidence that in cell culture GPI anchor-directed membrane association of PrP C is required for persistent PrP res propagation, implicating raft microdomains as a location for conversion.IMPORTANCE Mechanisms of prion propagation, and what makes them transmissible, are poorly understood. Glycosylphosphatidylinositol (GPI) membrane anchoring of the prion protein (PrP C ) directs it to specific regions of cell membranes called rafts. In order to test the importance of the raft environment on prion propagation, we developed a novel model for prion infection where cells expressing either GPI-anchored PrP C or transmembrane-anchored PrP C , which partitions it to a different location, were treated with infectious, misfolded forms of the prion protein, PrP res . We show that only GPI-anchored PrP C was able to convert to PrP res and able to serially propagate. The results strongly suggest that GPI anchoring and the localization of PrP C to rafts are crucial to the ability of PrP C to propagate as a prion.

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Strain‐Associated Variations in Abnormal PrP Trafficking of Sheep Scrapie

Cited by 35 publications

References 47 publications

Prion Protein with an Insertional Mutation Accumulates on Axonal and Dendritic Plasmalemma and Is Associated with Distinctive Ultrastructural Changes

Prion Protein with an Insertional Mutation Accumulates on Axonal and Dendritic Plasmalemma and Is Associated with Distinctive Ultrastructural Changes

Glycosylphosphatidylinositol Anchoring Directs the Assembly of Sup35NM Protein into Non-fibrillar, Membrane-bound Aggregates

PrP Knockout Cells Expressing Transmembrane PrP Resist Prion Infection

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