Prion Strain- and Species-Dependent Effects of Antiprion Molecules in Primary Neuronal Cultures

Cronier, Sabrina; Béringue, Vincent; Bellon, Alfredo; Peyrin, Jean‐Michel; Laude, Hubert

doi:10.1128/jvi.01502-07

Cited by 59 publications

(63 citation statements)

References 57 publications

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“…In the previous reports using prion-infected primary cultured neurons, antimitotics were used throughout the culture period to suppress the growth of astrocytes (Carimalo et al, 2005;Cronier et al, 2007;Hannaoui et al, 2013), and some of them confirmed the neuronal predominance with astrocytes at 4 -5 % of the cellular composition at an early stage (Cronier et al, 2004). However, antimitotics will be harmful to neurons depending on concentrations and the period of treatment.…”

Section: Discussionmentioning

confidence: 99%

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Tanaka

Fujiwara

Suzuki

et al. 2016

Journal of General Virology

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We established abnormal isoform of prion protein (PrP Sc )-specific double immunostaining using mAb 132, which recognizes aa 119-127 of the PrP molecule, and novel PrP Sc -specific mAb 8D5, which recognizes the N-terminal region of the PrP molecule. Using the PrP Sc -specific double immunostaining, we analysed PrP Sc in immortalized neuronal cell lines and primary cerebralneuronal cultures infected with prions. The PrP Sc -specific double immunostaining showed the existence of PrP Sc positive for both mAbs 132 and 8D5, as well as those positive only for either mAb 132 or mAb 8D5. This indicated that double immunostaining detects a greater number of PrP Sc species than single immunostaining. Double immunostaining revealed cell-type-dependent differences in PrP Sc staining patterns. In the 22 L prion strain-infected Neuro2a (N2a)-3 cells, a subclone of N2a neuroblastoma cell line, or GT1-7, a subclone of the GT1 hypothalamic neuronal cell line, granular PrP Sc stains were observed at the perinuclear regions and cytoplasm, whereas unique string-like PrP Sc stains were predominantly observed on the surface of the 22 L straininfected primary cerebral neurons. Only 14 % of PrP Sc in the 22 L strain-infected N2a-3 cells were positive for mAb 8D5, indicating that most of the PrP Sc in N2a-3 lack the N-terminal portion. In contrast, nearly half PrP Sc detected in the 22 L strain-infected primary cerebral neurons were positive for mAb 8D5, suggesting the abundance of full-length PrP Sc that possesses the Nterminal portion of PrP. Further analysis of prion-infected primary neurons using PrP Sc -specific immunostaining will reveal the neuron-specific mechanism for prion propagation.

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Section: Discussionmentioning

confidence: 99%

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Tanaka

Fujiwara

Suzuki

et al. 2016

Journal of General Virology

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“…Some transgenic lines have been established by a gene replacement method (for review [44]). PrP sequence identity between the transgenic host and donor usually lead to an enhanced susceptibility to TSE as compared to wild-type [58,127] mice. Moreover, in various instances, the TSE agent original properties, in particular PrP Sc molecular profile, appeared to be essentially maintained in the infected transgenic host (see also Sect.…”

Section: Influence Of the Prp Genementioning

confidence: 99%

Prion agent diversity and species barrier

2008

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-Mammalian prions are the infectious agents responsible for transmissible spongiform encephalopathies (TSE), a group of fatal, neurodegenerative diseases, affecting both domestic animals and humans. The most widely accepted view to date is that these agents lack a nucleic acid genome and consist primarily of PrP Sc , a misfolded, aggregated form of the host-encoded cellular prion protein (PrP C ) that propagates by autocatalytic conversion and accumulates mainly in the brain. The BSE epizooty, allied with the emergence of its human counterpart, variant CJD, has focused much attention on two characteristics that prions share with conventional infectious agents. First, the existence of multiple prion strains that impose, after inoculation in the same host, specific and stable phenotypic traits such as incubation period, molecular pattern of PrP Sc and neuropathology. Prion strains are thought to be enciphered within distinct PrP Sc conformers. Second, a transmission barrier exists that restricts the propagation of prions between different species. Here we discuss the possible situations resulting from the confrontation between species barrier and prion strain diversity, the molecular mechanisms involved and the potential of interspecies transmission of animal prions, including recently discovered forms of TSE in ruminants.prion / strain / misfolding / species barrier / PrP Table of Mammalian prion diseases or transmissible spongiform encephalopathies (TSE) form a group of related, invariably fatal neurodegenerative disorders of both animals and humans. The brain pathology consists of spongiosis, astrocytosis, neuronal loss and neural tissue from affected individuals contains an infectious agent, the prion, setting these diseases apart from other neurodegenerative diseases. Prion replication is thought to involve in essence the self-perpetuating conversion of the host-encoded cellular prion protein (PrP C ) into a misfolded form (PrP Sc ) that tends to aggregate and may be neurotoxic (for reviews [1,157]). Prion replication may also occur in lymphoid tissues but is there poorly if not pathogenic (for review [126], [133]). PrP C is a protein with two variably occupied glycosylation sites. It is attached at the outer of the plasma membrane by a glycosylphosphatidylinositol anchor. Its secondary structure is rich in alpha-helix and the protein is likely to be in a monomeric state in mild detergents. While its precise physiological function has not been assigned, PrP C is essential for prion replication and neurotoxicity to occur [57,129]. Upon infection, PrP C is refolded -without apparent post-translational modification -into beta-sheet -rich PrP Sc , initially in the presence of exogenous PrP Sc and then by an autocatalytic process. It leads to the formation of aggregates, sometimes of amyloid type, that can be differentiated from PrP C because of their partial resistance to protease digestion and of their insolubility into non-denaturing detergents [153]. Proteinase K, the most commonly used protease, di...

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“…Primary cultures of cerebellar granule neurons (CGN) were established as previously described (Cronier et al, 2007). Briefly, CGN cells were extracted from 5-or 6-d-old PrP knockout mice Zurich I (Büeler et al, 1992) by enzymatic and mechanical dissociation.…”

Section: Mouse Cerebellar Granule Neuronsmentioning

confidence: 99%

Neuron Dysfunction Is Induced by Prion Protein with an Insertional Mutation via a Fyn Kinase and Reversed by Sirtuin Activation inCaenorhabditis elegans

Bizat

Peyrin²,

Haı̈k³

et al. 2010

J. Neurosci.

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Although prion propagation is well understood, the signaling pathways activated by neurotoxic forms of prion protein (PrP) and those able to mitigate pathological phenotypes remain largely unknown. Here, we identify src-2, a Fyn-related kinase, as a gene required for human PrP with an insertional mutation to be neurotoxic in Caenorhabditis elegans, and the longevity modulator sir-2.1/SIRT1, a sirtuin deacetylase, as a modifier of prion neurotoxicity. The expression of octarepeat-expanded PrP in C. elegans mechanosensory neurons led to a progressive loss of response to touch without causing cell death, whereas wild-type PrP expression did not alter behavior. Transgenic PrP molecules showed expression at the plasma membrane, with protein clusters, partial resistance to proteinase K (PK), and protein insolubility detected for mutant PrP. Loss of function (LOF) of src-2 greatly reduced mutant PrP neurotoxicity without reducing PKresistant PrP levels. Increased sir-2.1 dosage reversed mutant PrP neurotoxicity, whereas sir-2.1 LOF showed aggravation, and these effects did not alter PK-resistant PrP. Resveratrol, a polyphenol known to act through sirtuins for neuroprotection, reversed mutant PrP neurotoxicity in a sir-2.1-dependent manner. Additionally, resveratrol reversed cell death caused by mutant PrP in cerebellar granule neurons from prnp-null mice. These results suggest that Fyn mediates mutant PrP neurotoxicity in addition to its role in cellular PrP signaling and reveal that sirtuin activation mitigates these neurotoxic effects. Sirtuin activators may thus have therapeutic potential to protect from prion neurotoxicity and its effects on intracellular signaling.

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Prion Strain- and Species-Dependent Effects of Antiprion Molecules in Primary Neuronal Cultures

Cited by 59 publications

References 57 publications

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Prion agent diversity and species barrier

Neuron Dysfunction Is Induced by Prion Protein with an Insertional Mutation via a Fyn Kinase and Reversed by Sirtuin Activation inCaenorhabditis elegans

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