Printer Friendly

Dual role of cellular prion protein in normal host and Alzheimer's disease.


Prion diseases or transmissible spongiform encephalopathies (TSEs) are fatal neurological disorders that include Creutzfeldt-Jakob disease (CJD) and kuru in humans, scrapie in sheep and goats, bovine spongiform encephalopathy (BSE) in cattle, and chronic wasting disease (CWD) in cervids. For the last 30 years, we and other Japanese groups have reported Japanese scrapie and BSE cases. (1-14) Hitherto, 36 cases of BSE (10, 13) have been reported in Japan (Fig. 1). Global elucidation of the etiology of kuru has lead to the discovery of pre-senile dementias, CJD and its variant--with basically similar cellular lesions--which are also transmissible and may be caused by a very similar and unconventional "virus" or prion. (15, 16) All the transmissible cerebral amyloidoses (TCA) are formed from the TCA amyloid precursor protein (PrP) specified on the short arm of chromosome 2 in humans and chromosome 20 in mice.

There are other slow infections of the central nervous systems in humans that are caused by conventional viruses, including measles virus, papovaviruses (JC virus and SV40-progressive multi-focal leukoencephalopathies or PML), rubella virus, cytomegalovirus, herpes simplex virus, adenovirus types 7 and 32, Russian spring-summer encephalitis (RSSE) viruses, and the human retroviruses (human T-cell lymphotropic viruses-1: HTLV1, and human immunodeficiency viruses: HIV) (Table 1). Recently, Notkins's group, our and other groups have demonstrated that human enteroviruses and reoviruses can be regarded as typical slow infections causing type-1 diabetes mellitus. (17-25) However, unlike these conventional viruses, the "unconventional viruses" of the sub acute spongiform encephalopathies are truly slow in their replication, with long doubling times. Moreover, these alone appear to be viruses that have made it necessary to alter our conceptions of the possible range of virus structure. The process of infection appears to be a seeding by a "virus", which is a nucleating agent inducing and the automatically accelerating the conformational transition in the amyloid subunit protein. In that process the host-specified precursor protein (cellular isoform of prion protein: Pr[P.sup.C]) is converted to an insoluble cross-[beta]-plated configuration [(scrapie isoform of prion protein: PrP.sup.Sc]) (26) (Table 2). Oligomers of microfilament of this subunit protein nucleate its own polymerization, crystallization, and precipitation as insoluble arrays of amyloid fibrils. Thus, proteolytic cleavage and conformational change of the precursor and oligomeric assembly of this structurally altered polypeptide produces a fibril amyloid enhancing factor. (27) with apparent infectious properties. In this report, the dual role of Pr[P.sup.C] is described in diseased and normal function in the compromised host. Additionally, the etiological role of Pr[P.sup.C] in the induction of Alzheimer's disease (AD) is discussed.

1. Pr[P.sup.C] is indispensable for neurological functions

1) Neuritogenesis by Pr[P.sup.C]. The mammalian Pr[P.sup.C] is a highly conserved glycoprotein localized in membrane lipid rafts and anchored to the cell surface by glycosylphpsphatidylinositol (GPI) (28, 29) Pr[P.sup.C] is located in many cell types, and is particularly abundant in neurons (30) Under certain conditions Pr[P.sup.C] may undergo conversion into a conformation-ally altered isoform (Pr[P.sup.Sc]) which is widely believed to be the pathogenic agent in prion disease or TSE. (31) Although much has been studied about the effects of Pr[P.sup.Sc] in prion diseases, the normal function of Pr[P.sup.C] is poorly understood as yet. Lehmann and our group have shown that Pr[P.sup.C] has [alpha]- and [beta]-cleavage site during normal processing and host translational modifications. (32)

Recent experimental evidence showing that Pr[P.sup.C] interacting with [beta]1 integrin controls focal adhesion and turnover of the actin microfilaments in neurons substantiates a role for Pr[P.sup.C] in neurito-genesis. Several reports show that Pr[P.sup.C] participates in trans-membrane signaling processes associated with hematopoietic stem cell replication and neuronal differentiation. (33-35) Integrins are well known in autophagy. Remarkably, during neuronal differentiation, the downregulation of Rho kinase activity is necessary for neurite sprouting. (33) Abundant expression of Pr[P.sup.C] has been detected during mouse embryogenesis in association with the developing mouse nervous system. (36) In the developing mouse brain, undifferentiated neural progenitor cells in the mitotically active ventricular zone do not express Pr[P.sup.C]. In contrast, post-mitotic neurons express high levels of Pr[P.sup.C] after their last mitosis in the neuro-epithelium as they migrate towards marginal layers and differentiate. (36, 37) Thus, Pr[P.sup.C] may be expressed exclusively in differentiated neurons.

Studies in vitro have shown that expression of Pr[P.sup.C] is positively correlated with differentiation of multipotent neuronal precursors into mature neurons. (37) In addition, treatment of embryonic hippocampal neurons with recombinant Pr[P.sup.C] enhance neurite outgrowth and survival. (38) The distribution of Pr[P.sup.C] in the developing nervous systems of cattle, (39) mice, (36) and humans(40) suggest that Pr[P.sup.C] plays a functional role in neural development. While Pr[P.sup.C]-knockout ([Prnp.sup.-/-]) displays no overt in neural phenotype, (41) numerous subtle phenotypes have been reported, (37) including reduction in the number of neural precursor cells in developing mouse embryo. (35) Other studies have shown that Pr[P.sup.C] induced neuritogenesis in embryonic hippocampal neurons cultured in vitro. (38, 42) Pr[P.sup.C] interacts with stress inducible protein 1 (STI1, 43) which is a heat-shock protein. (44) The interaction of Pr[P.sup.C] with STI1 not only activates cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) to transduce a survival signal but also induces phosphorylation/activation of the mitogen-activated protein kinase to promote neuritogenesis. (42) Elucidation of the signaling pathways through which Pr[P.sup.C] influences neurogenesis, along with an understanding of the pathways involved in other key cellular processes, (42) may provide insight into how Pr[P.sup.C] misfolding leads to devastating neurodegenerative diseases.

2) Neuroprotection by Pr[P.sup.C]. In this chapter we describe how our group has spent much effort to understand the normal function of Pr[P.sup.C] using [Prnp.sup.-/-] mice. Interestingly we observed that Pr[P.sup.C] worked in an in vitro system favorably provide neuroprotection for mice under the oxidative stress or virus infection by suppressing the apoptosis. In this chapter and Chapter 4, we show the same neuroprotective role of Pr[P.sup.C] in an in vivo mouse system. The most commonly observed function on Pr[P.sup.C] is copper-binding. Previous and our studies by our laboratories and others have demonstrated that the octapeptide-repeat region of Pr[P.sup.C] binds with [Cu.sup.2+] within the physiological concentration range. (45-47) Furthermore, Pr[P.sup.C] displays a functional role in normal brain metabolism of copper. (48) Besides binding with [Cu.sup.2+] at the synapse, Pr[P.sup.C] serves as [Cu.sup.2+] buffer as well. (49) Over expression of Pr[P.sup.C] increases [Cu.sup.2+] uptake into cells, (50) while Pr[P.sup.C] knockout ([Prnp.sup.-/-]) mice show a lower synaptosomal [Cu.sup.2+] concentration than normal mice. (49) However, [Cu.sup.2+] rapidly and reversibly stimulates the internalization of Pr[P.sup.C] during Pr[P.sup.C] endocytosis. (51, 52) Through binding with [Cu.sup.2+,] Pr[P.sup.C] displays super oxide dismutase (SOD) activity in vitro. (50, 53-57) Intriguingly, treatment with copper chelator cuprizone induces TSE-like spongiform degeneration.(58) Therefore, [Cu.sup.2+] metabolism appears to play an important role in not only PrP function but also the pathogenesis of prion diseases.

Pr[P.sup.C] may act as an anti-apoptotic agent by blocking some of the internal or environmental factors that initiate apoptosis. (59, 60) Mature Pr[P.sup.C] trends to localize in lipid rafts of cells. (30) As lipid rafts are membrane structures specialized in signaling, a potential role of Pr[P.sup.C] in signal trans diction may be anticipated. A[beta] on the raft is the most important interacting protein among them (see the next chapter: section 3). Such a signaling platform or signal some is based on Pr[P.sup.C]. Pr[P.sup.C] localizes to cholesterol- and sphingolipid-rich, detergent resistant lipid rafts due to the saturated acryl chains in its GPI anchor and to an N-terminal targeting signal interacting with heparin sulfate proteoglycan, glypican-1. Pr[P.sup.C] has been proposed as a key scaffolding protein for dynamic assembly of cell surface signaling modules, and Pr[P.sup.C], along with the micro domain-forming flotillin, or caveolin proteins, may lead to the local assembly of membrane protein complexes at sites involved in cellular communication, such as cell-cell contacts, focal adhesions, the T-cell cap, and synapses.

In addition, a phosphorylating function of Pr[P.sup.C], mediated by caveolin-1 to indirectly increase Fyn (a member of the Src family of tyrosine kinase) phosphorylation, governs the downstream production of NADPH-oxidize-dependent reactive oxygen species and activation of the extra-cellular regulated kinase 1/2 has been demonstrated. (30, 61) Pr[P.sup.C] interacts with normal phosphoprotein synapsin Ib and cytoplasmic adaptor protein Grb2 without being deciphered with prion interactor Pint1. (62) Bovine PrP strongly interacts with the catalytic [alpha]/[alpha]' subunit of protein kinase CK2 to increase the phosphotransferase activity of CK2, thus leading to the phosphorylation of calmodulin. (63)

Upon engagements with Pr[P.sup.C] binding peptides and certain antibodies, Pr[P.sup.C] transduces neuroprotective signals that affect the sensitivity to induce cell death. (64) Linden et al. have suggested that the engagement of Pr[P.sup.C] transduces neuroprotective signals through a cAMP/PKA-dependent pathway.

According to their theory, Pr[P.sup.C] may function as a trophic receptor, where the activation of which leads to a neuroprotective state. (64) In addition, Pr[P.sup.C] binds with extracellular matrix laminin to promote genesis and maintenance of neurites. We have shown that PI3K is activated by Pr[P.sup.C] through the intercellular transportation of copper molecules. (65) In fact, a study has discovered Pr[P.sup.C] to induce self-renewal of longterm populating hematopoietic stem cells. (34) Furthermore, another study has revealed that Pr[P.sup.C] is expressed on the multi-potent neural precursors and mature neurons without being detected in glia, suggesting that Pr[P.sup.C] plays an important role in neural differentiation. (39) Therefore, the interaction between Pr[P.sup.C] and various signal transudation molecules speaks well for important functions (such as differentiation and cell survival) in the living system (Fig. 2).

A stress-protective activity has also been assigned to Pr[P.sup.C] based on results with primary neuronal cultures. Neuronal cells derived from [Prnp.sup.-/-] mice are more sensitive to oxidative stress and serum deprivation than wild-type cells. Moreover, after ischemic brain injury, [Prnp.sup.-/-] mice reveal enlarged infarct volumes. (66) In our studies [Prnp.sup.-/-] cell line (HpL3-4), immortalized from hippocampal neuronal precursor cells in Rikn [Prnp.sup.-/-] mice (Fig. 3), is sensitive to serum deprivation-induced apoptosis, although it is activated and/or survives with Pr[P.sup.C] expression. (53) The same effect was observed in another Prnp-deficient cell line (NpL2), immortalized from hippocampal neuronal cells in Zrch I [Prnp.sup.-/-] mice. (57) Over expression of B-cell lymphoma 2 protein (Bcl-2) in this cell line reveals a functional relation of Pr[P.sup.C] with Bcl-2 in the anti-apoptotic pathway. (53) Martin et al. have shown that prevention of cell death in cultured retinal explants from neonatal rats and mice induced by anisomycin (a protein synthesis inhibitor) unfurls the effect associated with Pr[P.sup.C]-STI1 interactions. (43) Li and our group show that the production of another type of heat-shock protein (Hsp 70) is enhanced when PrP levels elevate during hyperglycemia. (67)

We have demonstrated that the inhibition of apoptosis through STI1 is mediated by Pr[P.sup.C]-dependent SOD activation. (68) This neuroprotective role of Pr[P.sup.C] has been linked to cell signaling events. The interaction of Pr[P.sup.C] with the STI1 generates neuro-protective signals that rescue cells from apoptosis (Fig. 4). The functional role of STI1 and Pr[P.sup.C] have been confirmed in both murine and bovine systems. (69)

The late onset of severe ataxia and loss of cerebellar Purkinje cells in several [Prnp.sup.-/-] mouse lines (70-72) from several laboratories suggest the lack of cerebellar protection by Pr[P.sup.C] in these mice. Interestingly, deposition of Pr[P.sup.Sc] has been located in the deep cerebellar nuclei (DCN) of scrapie-infected sheep. (73) Future studies with a microarray analysis (74) by our group applied in eye-blink-conditioning of mice may provide insight into understanding the normal function of Pr[P.sup.C] in the DCN of the cerebellum.

A loss of Pr[P.sup.C] function could be implicated in the pathogenesis of prion diseases and Pr[P.sup.C]-dependent pathways might be involved in neurotoxic signaling. For example, in vivo crosslinking of Pr[P.sup.C] by antibodies triggered neuronal apoptosis (75) and Pr[P.sup.C]-dependent receptors were postulated to explain the neurotoxic effect of a PrP mutant lacking the hydrophobic domain (see next sections 2 and 3). (76) Still available data indicated that Pr[P.sup.C] is a critical element of the network that controls the sensitivity to programmed cell death in both the nervous system and several other cell types. The outcome of the engagement of Pr[P.sup.C] on either cell death or survival is likely dictated by its available ligands. These ligands in fact determine the array of intervening signaling pathways.

Taken together, Pr[P.sup.C] is functionally involved in copper metabolism, signal transduction, neuroprotection, and cell maturation. Despite these putative roles, mice null for Pr[P.sup.C] display no consistent phenotype apart from complete resistance to TSE infection. (41, 77) Further searches for Pr[P.sup.C]-interaction molecules using [Prnp.sup.-/-] mice and various types of [Prnp.sup.-/-] cell lines under various conditions may elucidate the Pr[P.sup.C] functions.

3) Enhanced synaptic plasticity by Pr[P.sup.C]. In Rikn [Prnp.sup.-/-] mice, Kim et al. in our group have observed pathological alterations and some physiological dysfunctions in the olfactory bulb (OB). (78) Recently, Le Pichon et al. have proposed that electrophysiological alterations at the dendrodendritic synapse in the OB could underlie the behavior phenotypes. (79) In detail, the cookie finding phenotype was manifested in three [Prnp.sup.-/-] lines (Zrch I [Prnp.sup.-/-] mice, (41) Ngsk [Prnp.sup.-/-] mice, (72) Npu [Prnp.sup.-/-] mice, (80) on alternate genetic backgrounds, indicating strong evidence of its dependence on Pr[P.sup.C] rather than other genetic factors. [Prnp.sup.-/-] mice also displayed altered behavior in the habituation-disambiguation task, suggesting the phenotype is most likely to be olfactory-specific. (78, 79) [Prnp.sup.-/-] mice exhibited wide spread alterations of oscillatory activity in the OB as well as altered paired-pulse plasticity at the dendrodendritic synapse. Importantly, both the behavioral and electrophysiological phenotypes could be rescued by neuronal Pr[P.sup.C] expression. (79) These data suggest a critical role for Pr[P.sup.C] in the normal processing of sensory information by the olfactory system.

Kim et al. and Le Pichon et al. employed a so-called "cookie finding task", a test of broad olfactory acuity, to analyze a battery of mice including PrP knockout from multiple genetic backgrounds and transgenic mice in which Prnp expression was driven by cell type-specific promoters. (78, 79) [Prnp.sup.-/-] mice exhibited impaired behavior that was rescued in transgenic mice expressing Pr[P.sup.C] specifically in neurons but not in mice expressing only extra neuronal Pr[P.sup.C]. [Prnp.sup.-/-] mice displayed altered behavior in an additional olfactory test (habituation-dishabituation) which was also rescued by transgenic neuronal PrP expression suggesting that the phenotype was olfactory-specific. Additionally, the odor evoked electrophysiological properties of the OB of [Prnp.sup.-/-] mice have been studied. (79) In these mice, alterations in the patterns of oscillatory activity in the OB were detected. The plasticity of dendrodendritic synaptic transmission was altered between the granule cells and mitral cells.

Disruption was observed in local field potential (LFP) oscillation and in the plasticity of the dendrodendritic synapse, either, or both, of which could contribute to the behavioral phenotype in [Prnp.sup.-/-] mice. Oscillatory LFPs may act to organize information flow within the olfactory system (81, 82) by constraining the timing of mitral cell action potentials. In addition, gamma oscillations are specifically implicated in behavioral performance in olfactory tasks. (83, 84) Therefore, alterations in oscillatory timing during odor exposure may perturb OB output to higher centers by disrupting how information is packaged within a breathing cycle.

Altering the dendrodendritic synapse may have multiple functional consequences. This synapse may mediate lateral inhibition between ensembles of mitral cells, and be critical for olfactory discrimination. (85, 86) Additionally, because granule cells receive convergent information onto their proximal dendritic arbor from multiple higher brain areas, (87) disruption of the dendrodendritic synapse may alter the transmission of centrifugal modulation of OB mitral cells.

High-frequency oscillations in the OB are shown to result from the rapid and reciprocal interactions between granule and mitral cells across the dendrodendritic synapse in vitro. (88) Therefore, Le Pichon's observation could imply that increased facilitation of mitral cell inhibitory postsynaptic potential (IPSP) following repetitive spiking decreases the dynamic range and increases the duration of gamma oscillations across the boundaries of breath. Although both oscillatory and synaptic effects could be reversed by neuronal Pr[P.sup.C] expression, they cannot claim a causal link between these findings. Mitral cells receive facilitated inhibition in [Prnp.sup.-/-] mice. Future work should determine the precise synaptic localization of the Pr[P.sup.C] protein as well as its biochemical interactions with synaptic machinery.

At least two definite molecular interactions of Pr[P.sup.C] with hippocampal cell surface proteins, STI1 and laminin, can mediate the effect of Pr[P.sup.C] on memory consolidation. (89) Moreover, it is likely that Pr[P.sup.C] modulates memory retention through both these interactions. Further support of the hypothesis that Pr[P.sup.C] plays important roles in memory and cognition is found in humans. (89) The presence of Val at codon 129 of PRNP (human prion protein gene) in at least one allele is associated with worsened cognitive performance in elderly subjects, with early cognitive decline.89) Conversely, healthy young adults expressing Met at codon 129 in at least one allele exhibit better long-term memory than those with Val in this codon, although short-term memory was unaffected. Thus, polymorphism at codon 129, a site that is highly important for protein structure, seems to be strongly related to cognitive performance. (89)

Another report shows the potential impact of the temporary disruption of the cerebellar circuit during the time window critical for sensor motor development in Zrch I [Prnp.sup.-/-] mice. (90) In that investigation, the authors considered the effect of PrP gene knock-out on cerebellar neural circuits, which show intense PrP expression during development and selective affinity for PrP. Zrch I [Prnp.sup.-/-] mice showed low performance in the accelerating rotarod and runaway test and the functioning of 40% of granule cells was abnormal. This phenomenon may have a reflection in some of the late motor, cognitive, and emotional abnormalities in Zrch I [Prnp.sup.-/-] mice. (90) Their results suggest that PrP plays an important role in granule cell development to eventually regulate cerebellar network and motor control.

4) Myelination in peripheral nerves by Pr[P.sup.C]. A late-onset peripheral neuropathy has been identified in Pr[P.sup.C]-deficient Ngsk [Prnp.sup.-/-] and Zrch I Prnp!!! mice (41, 72, 91) (Fig. 4). This indicates that Pr[P.sup.C] might have a role in peripheral neuropathies.

At 60 weeks of age, all investigated Zrch I [Prnp.sup.-/-] mice (n = 52) showed chronic demyelinating polyneuropathy (CDP). (92) CDP was 100% penetrating and conspicuous in all investigated peripheral nerves (sciatic and trigeminal nerves, dorsal and ventral spinal roots) (Fig. 4). Besides, CDP was associated with another 2 independently targeted Prnp knockout mouse lines, GFP [Prnp.sup.-/-] mice (93) and Npu [Prnp.sup.-/-] mice. (80)

Zrch I [Prnp.sup.-/-] and Npu [Prnp.sup.-/-] mice suffered from CDP despite normal expression of Doppel (Dpl, 70) indicating that Dpl regulation did not cause polyneuropathy. CDP was present in mice lacking both Prnp and Prnd (the gene for Dpl, 94) but absent from mice selectively lacking Prnd. (95) Therefore, Dpl is not required for the maintenance of peripheral nerves. Pr[P.sup.C] might interact with myelin component directly or through other axonal proteins. Certain reported Pr[P.sup.C] interacting proteins have roles in homeostasis, (96) and represent possible candidates for mediation of its myelinotrophic effects. The octapeptide repeat region was not required for myelin maintenance, whereas mice PrP lacking central domain (aa 94-134) developed CDP. (97) It is worthy to note that the hydrophobic core, but not the charge cluster (C[C.sub.2]), of this central Pr[P.sup.C] domain was essential for peripheral myelin maintenance.

Our and many other recent reports have shown that Pr[P.sup.C] undergoes regulated proteolysis in late secretory compartments. (32, 98-100) Bremer et al. (92) have observed an association between the presence of CDP and lack of C1 fragment in the sciatic nerves. All Pr[P.sup.C] mutants in which CDP was rescued produced abundant C1. (92) CDP was prevented by Pr[P.sup.C] variants that undergo proteolytic amino-proximal cleavage, (32) but not by variants that undergo non-permissive cleavage, including secreted Pr[P.sup.C] lacking its glycolipid membrane anchor. Cleavage of Pr[P.sup.C] appeared therefore to be linked to its myelinotrophic function. This conjuncture might also explain the requirement for membrane anchoring of Pr[P.sup.C] uncovered in GPI-deficient PrP transgenic ([]GPI-PrP) mice, (101) as anchorless Pr[P.sup.C] did not undergo regulated proteolysis. These results indicate that neuronal expression and regulated proteolysis of Pr[P.sup.C] are essential for myelin maintenance. (92)

Nervous myelin degeneration in optic nerves, corpus callosum or spinal cords was not detected in 60-week-old Zrch I [Prnp.sup.-/-] mice. (92) Nevertheless subliminal myelin pathologies might extend to central myelin in Zrch I [Prnp.sup.-/-] mice, (102) and transgenic mice expressing toxic Pr[P.sup.C] show both peripheral and central myelinopathies. (97, 103) Pr[P.sup.C] deficiency has been reported to affect synaptic function, (104, 105) however, the amplitudes of foot muscle compound action potentials following distal stimulation are not significantly altered in 53-week-old Zrch I [Prnp.sup.-/-] mice; arguing against an important synaptic defect in the neuromuscular synaptic junction.

It has been suggested that Pr[P.sup.C] has various roles in immunity, (106) and lymphocytes are important in mouse models of hereditary demyelinating neuropathies. As the CDP in our mutant mice was not modulated by Rag1 removal, lymphocytes were not involved in its pathogenesis. The combined results of restricting expression of Pr[P.sup.C] of neurons and of selectively depleting Pr[P.sup.C] from neurons indicate that the expression of Pr[P.sup.C] by the neuron is essential for the long-term integrity of peripheral myelin sheaths. (92) Not only was the trophic function of Pr[P.sup.C] exerted in trans, but also correlated with the proteolytic processing of in diverse transgenic mouse models. These findings identify Pr[P.sup.C] as a critical messenger of trans-cellular axomyelinic communication, and indicate that regulated proteolysis of axonal Pr[P.sup.C] might expose domains that interact with Schwann cell receptors. Recently, Aguzzi's group has reported that Pr[P.sup.C] promotes myelin homeostasis through flexible tail (23-50)-mediated Gpr126 agonism, (107) and Gpr126 is crucial for peripheral nerve development. [Gpr126.sup.-/-] mice exhibit drastic hypomyelination. (107) Clarifying the molecular basis of these phenomena might lead to better understanding of peripheral neuropathies, particularly those with late-onset, and might help to uncover new therapeutic targets.

2. Pr[P.sup.C] mediates toxic signaling by Pr[P.sup.Sc]

Mice with prion disease show Pr[P.sup.Sc] accumulation and develop extensive neurodegeneration, in contrast to mouse models of AD or Parkinson's disease, where neuronal loss is rare. Therefore, prion-infected mice allow access to mechanisms linking protein misfolding to neuronal death. Studies of both neuronal and nonneuronal cells substantiate the coupling of Pr[P.sup.C] to signaling effectors involved in cell survival, redox equilibrium and homeostasis (e.g., ERK1/2, NADPH oxidase, cyclic AMP-responsive element binding protein (CREB) transcription factor and metalloproteinases) (Fig. 2). According to these studies, Pr[P.sup.C] functions as a dynamic cell surface platform for assembly of signaling molecules. However, the sequence of cellular and molecular events that leads to neuronal cell demise in TSEs remains obscure. At present, we envision that neuronal cell death results from several parallel, interacting or sequential pathways involving protein processing and proteasome dysfunction, oxidase stress, apoptosis, and autophagy.

The fact that Pr[P.sup.Sc] levels alone cannot serve as a marker for tissue infectivity suggests that it may be useful to adapt current protocols of prion detection in tissues, since they are so far largely based on the detection of bona fide Pr[P.sup.Sc]. (108) Mallucci's group has previously shown rescue of neuronal loss and reversal of early cognitive and morphological changes in prion-infected mice by depleting Pr[P.sup.C] in neurons, preventing prion replication and abrogating neurotoxicity. (109) Recently, the same group has shown that Pr[P.sup.Sc] replication causes sustained unfolded protein response

(UPR) induction with persistent, deleterious expression of eLF2a-P in prion disease. (110) The resulting chronic blockade of protein synthesis leads to synaptic failure, spongiosis, and neuronal loss. Promoting eLF2a-P dephosphorylation rescues vital translation rates and is thereby neuroprotective, whereas preventing this further reduces translation and enhances neurotoxicity. The data support the development of generic proteostatic approaches to therapy in prion diseases. (111),112) The unfolded Pr[P.sup.C] response works as a protective cellular mechanism triggered by rising levels of misfolded Pr[P.sup.Sc] protein. (110)

In another study, expression of Pr[P.sup.C] in neuronal cells is required to mediate neurotoxic effects of Pr[P.sup.Sc], (101) which might elicit a deadly signal through a Pr[P.sup.C]-dependent signaling pathway. Spontaneous neurodegeneration in transgenic mice expressing a Pr[P.sup.C] mutant without the N-terminal ER-targeting sequence indicated the toxic potential of Pr[P.sup.C] when located in the cytosolic compartment (cytoPrP). (113) Toxicity of cytoPrP seems to be dependent on its association with cellular membranes (114) and its binding to Bcl-2, an antiapoptotic protein present at the cytosolic side of ER and mitochondrial membranes. (115) Might the toxic potential of misfolded Pr[P.sup.C] in the cytosol be relevant to the pathogenesis of prion diseases? The most recent finding has revealed an impairment of the ubiquitin-proteasome system (UPS) in prion-infected mice. In conjunction with in vitro and cell culture approaches, it has been proposed that prion neurotoxicity is linked to Pr[P.sup.Sc] oligomers, which translocate to the cytosol and inhibit the UPS. (116) Another study shows that a reduction in autophagy combined with endosomal/ lysosomal dysfunction has indeed been proposed for the development of prion diseases. (117)

3. Role of Pr[P.sup.C] in AD

Pr[P.sup.C] is anchored to the cell surface by GPI. Since the 1980s, PrP-related studies have mainly focused on the mechanisms by which Pr[P.sup.C] is converted into Pr[P.sup.Sc] that is responsible for transmissible spongiform encephalopathies (TSE) and causes fatal neurodegeneration and aggregation. Concomitantly, the physiology of Pr[P.sup.C] has also been studied, including the role in the cellular trafficking of copper ions, hippocampal morphology, cognition, oxidative stress and apoptosis (118) (Fig. 5). Depending on cell context or physiological conditions, Pr[P.sup.C] is beneficial or harmful to the cell. Many studies have indicated that Pr[P.sup.C] can rescue neurons from various stressful situations. (53)'(119) Evidence from Prnp-knockout mice studies has provided information regarding subtle phenotypes (e.g., mild neuropathological, cognitive and behavioral deficits, 120) and enhanced brain injury after ischemia. (66)'(121) In addition, the N-terminal region of Pr[P.sup.C] displays a neuroprotective function. (122) Conversely, Pr[P.sup.C] also tends to function as a neurotoxic protein. (123)

Pr[P.sup.C] expression is indispensable for prion-induced neurotoxicity to develop (124) implying Pr[P.sup.C] could be a receptor for prions to trigger detrimental signaling. Strittmatter has reported that Pr[P.sup.C] transduces the synaptic cytotoxicity of amyloid-[beta](A[beta]) oligomers in vitro (125) and in A[beta]-transgenic mice. (126) Moreover, different anti-PrP antibodies or their antigen-binding fragments that disrupt the PrP-A[beta] interaction are able to block the A[beta]-mediated disruption of synaptic plasticity. These findings are important because they suggest the involvement of Pr[P.sup.C] in AD pathogenesis. However, others have found that the absence of Pr[P.sup.C] does not prevent deficits in hippocampal-dependent behavioral tests upon intracerebral A[BETA] injection. (127) It has been suggested that variations in copper availability may contribute to these discrepancies. (128)

Pr[P.sup.C] seems to regulate the [beta]-secretase cleavage of amyloid precursor protein, thereby regulating the production of A[beta]. (129) In addition, [beta]-secretase regulates the cleavage of Pr[P.sup.C], regulating an N-terminal fragment with neuroprotective activity. (122, 130) Pr[P.sup.C] also binds to transmembrane proteins such as the 67kDa laminin receptor (131) neural cell adhesion molecules, (131)'(132) G protein-coupled serotonergic receptors, (64) and low density lipoprotein receptor-related protein 1 (LPR1),133),134) which are able to promote intracellular signaling-mediated neuronal adhesion and differentiation as well as Pr[P.sup.C] internalization.

Remarkably, Pr[P.sup.C] functions as a receptor or co-receptor for extracellular matrix proteins such as laminin (65) and vitronectin, 135) as well as STI1 (136) which has been repeatedly found by our group. These data suggest that GPI-anchored Pr[P.sup.C] is a potential scaffold receptor in a multi-protein, cell surface, and signaling complex that may serve as the basis for the multiple neuronal functions ascribed to Pr[P.sup.C]. (89, 137)

Pr[P.sup.C] has been identified to bind A[BETA] oligomers (A[BETA]O), but not monomers or fibrils, with high affinity and to selectively interact with high molecular mass assembles of A[BETA]O in AD but not control brains. (30) Pr[P.sup.C] is responsible for A[BETA]O-mediated inhibition of long-term potentiation (LTP) in hippocampal slices and is also required for the manifestation of memory impairment in an AD mouse model. A[BETA]O-binding to Pr[P.sup.C] leads to activation of Fyn kinase. In addition, the A[BETA]O activation of Fyn leads to tau phosphorylation. Both metabotropic glutamate receptor 5 (mGluR5) and LPR1 have been identified as co-receptors required for the Pr[P.sup.C]bound A[BETA]O to activate Fyn.30) Fyn kinase phosphorylates N-methyl-D-aspartate receptor (NMDAR) and tau. Eventually NMDAR and tau (pTyr18) induce synaptic impairment and neurodegeneration (Fig. 6).

Recently, A[[BETA].SUB.42], which is associated with neuro-degeneration in AD, has also been reported to act as a ligand of Pr[P.sup.C]. (138) However, the physiological role of Pr[P.sup.C] as an A[[BETA].SUB.42]-binding protein is not clear. Actually, Jung and our group have demonstrated that Pr[P.sup.C] is critical in A[[BETA].SUB.42]-mediated autophagy in neurons. (138) The interaction of Pr[P.sup.C] with Beclin (BECN)1 facilitates the localization of BECN1 into lipid rafts and thus allows the activation of phosphatidylinositol 3-kinase (catalytic subunit type-3 or PI3KC3) complex in response to A[[BETA].SUB.42], showing a beneficial role of Pr[P.sup.C] as a positive regulator of the BECN1-PI3KC3 complex in lipid rafts.

4. Pr[P.sup.C] and viral or bacterial infection

Microglia is one of the major cell types that produce NO, which in fact plays a role in active cellular protection. Keshet et al. and our group have reported that NOS activity is significantly reduced in adult Zurich I [Prnp.sup.-/-] mice (age > 100 days), whereas NOS activity in young Zrch I [Prnp.sup.-/-] mice ([less than or equal to] 30-day-old) is similar to that of wild-type [Prnp.sup.+/+] mice. (139)'(140) To further elucidates the role of Pr[P.sup.C], we have infected wild type prion protein gene ([Prnp.sup.+/+]) and Zrch I or Rikn [Prnp.sup.-/-] mice with encephalomyocarditis virus (EMCV)-B via an intra cranial route. (140) With regard to the inflammatory response in EMCV-B-infected mouse brains, mild to severe inflammatory changes have been observed only in 15-week-old but not in 6-week-old [Prnp.sup.+/+] mice. There is histopathological evidence of suppressed microglial response in [Prnp.sup.-/-] mice, whereas these are more enlarged apoptotic brain lesions after EMCV infection in [Prnp.sup.-/-] mice. (140)

Decreases in the activities of certain enzymes (superoxide dismutase and catalase) have been observed in the brains of Zrch I [Prnp.sup.-/-] mice.(60,60,141) This enzyme-related deficiency may induce an intracellular oxidative state in mouse brain cells. Increased lipoperoxidation, a phenomenon reported in other neurodegenerative diseases such as Alzheimer's diseases and epilepsy, (142, 143) has been observed in such mice. In the absence of Pr[P.sup.C], increased oxidation of lipids and protein has been observed in the brain. (144) These findings suggest the physiological function of Pr[P.sup.C] is related to the cellular antioxidant defense system: viz., loss of the antioxidant defense system in [Prnp.sup.-/-] mice may contribute to more severe lesions in the brain after EMCV infection.

A murine model of streptococcal sepsis that mimics many aspects of pathogenesis and immunity in the human disease has been investigated. (145) Accordingly, they found that although Zrch I Prnp!!! mice had impaired ability to clear Streptococcus pyogens at the inoculation site in the thigh muscle (an event that is largely neutrophil-dependent), these mice were protected from bacteremia and sepsis. This protection was accompanied by a standard decrease in most serum cytokine concentrations, the exceptions being IL-9 and interferon-a. It is tempting to speculate that the inability of Zrch I [Prnp.sup.-/-] mice to mount a full-blown cytokine response may confer some protection from full-scale bacterial sepsis. Their findings are noteworthy in the context of the neutrophil response to zymosan-induced peritonitis in Zrch I [Prnp.sup.-/-] mice, (146) which may be caused by deficient IL-17 production leading to inadequate neutrophil migration.

Recently, it has been reported that mitochondria control the activation of NLRP3 (nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3) inflammasome, and that the inflammasome activation is negatively regulated by autophagy and positively regulated by reactive oxygen species (ROS).14[degrees] Pr[P.sup.C] has SOD activity and protects neurons from oxidative stress. Antioxidant Pr[P.sup.C] may contribute to suppressing inflamma some activation. The relationship between Pr[P.sup.C] and inflammasome remains to be fully elucidated.

DOI: 10.2183/ pjab.93.010


The author thanks Dr. Akikazu Sakudo, Associate Professor, Ryukyu University, for continuous support and help for this slow virus projects. Thanks are due to Dr. Makoto Haritani, Member of Research Center for Food Safety, University of Tokyo, and Dr. Katsuaki Sugiura, Professor of University of Tokyo, for preparing and reading this manuscript. This study was supported by grants-in-aid from the Ministry of Health, Labor and Welfare, Japan.


(1) Onodera, T., Haritani, N., Narita, M. and Nakagawa, M. (1990) Epidemiology of ovine scrapie in Japan. Jpn. Agric. Res. Q. 24, 216-218.

(2) Onodera, T., Ikeda, T., Muramatsu, Y. and Shinagawa, M. (1993) Isolation of scrapie agent from the placenta of sheep with natural scrapie in Japan. Microbiol. Immunol. 37, 311-316.

(3) Maas, E., Geissen, M., Groschup, M.H., Rost, R., Onodera, T. and Schatzl, H. (2007) Scrapie infection of prion protein-deficient cell line upon ectopic expression of mutant prion protein. J. Biol. Chem. 282, 18702-18710.

(4) Marshall, K.E., Hughson, A., Vascellari, S., Priola, Sakudo, A., Onodera, T. and Baron, G.S. (2016) PrP knockout cells expressing transmembrane PrP resist prion infection. J. Virol. 91, e01686-16.

(5) Onodera, T. and Hayashi, T. (1994) Diversity of clinical signs in natural scrapie cases occurring in Japan. Jpn. Agric. Res. Q. 28, 59-61.

(6) Onodera, T. and Saeki, K. (2000) Japanese scrapie cases. Jpn. J. Infect. Dis. 53, 56-61.

(7) Hosokawa, T., Tuchiya, K., Sato, I., Takeyama, N., Ueda, S., Tagawa, Y., Kimura, K.M., Nakamura, 1., Wu, G., Sakudo, A., Casalone, C., Mazza, M., Caramelli, M., Takahashi, H., Sata, T., Sugiura, K., Baj, A., Toniolo, A. and Onodera, T. (2008) A monoclonal antibody (1D12) defines novel distribution patterns of prion protein (PrP) as granule in nucleus. Biochem. Biophys. Res. Commun. 366, 657-663.

(8) Uraki, R., Sakudo, A., Michibata, K., Ano, Y., Kono, J., Yukawa, M. and Onodera, T. (2011) Blocking of FcR suppresses the intestinal invasion of scrapie agents. PLoS One 6, e17928.

(9) Sugiura, K., Yokoyama, T., Kumagai, S. and Onodera, T. (2003) A model to assess the risk of the introduction into Japan of bovine spongiform encephalopathy agent through imported animals, meat and meat-and-bone meal. Rev. Sci. Tech. 22, 777-794.

10) Onodera, T. and Kim, C.K. (2006) BSE situation and establishment of Food Safety Commission in Japan. J. Vet. Sci. 7, 1-11.

(11) Onodera, T., Sakudo, A., Wu, G. and Saeki, K. (2006) Bovine spongiform encephalopathy in Japan: history and recent studies on oxidative stress in prion diseses. Microbiol. Immunol. 50, 565-578.

(12) Sugiura, K. and Onodera, T. (2008) Cattle trace ability system in Japan for bovine spongiform encephalopathy. Vet. Ital. 44, 519-526.

(13) Sugiura, K., Onodera, T. and Bradley, R. (2009) Epidemiological features of the bovine spongiform encephalopathy epidemic in Japan. Rev. Sci. Tech. 28, 945-956.

(14) Kusama, T., Hibino, H., Onodera, T. and Sugiura, K. (2009) Animal feed controls implemented in Japan for the eradication of bovine spongiform encephalopathy. Vet. Ital. 45, 287-295.

(15) Gadjusek, D.C. (1977) Unconventional viruses and the origin and disappearance of kuru. Science 197, 943-960.

(16) Prusiner, S.B. (1991) Molecular biology of prion diseases. Science 252, 1515-1522.

(17) Thomas, H. (2016) Diabetes: Enterovirus dysregulates islet miRNAs. Nat. Rev. Endocrinol. 12, 2.

(18) Yoon, J.W., Onodera, T. and Jenson, A.B. (1978) Virus-induced diabetes mellitus. XI. Replication of coxsackie B3 in human pancreatic beta cells in culture. Diabetes 27, 778-781.

(19) Yoon, J.W., Austin, M., Onodera, T. and Notkins, A.L. (1979) Virus-Induced diabetes mellitus: Isolation of virus from the pancreas of child with diabetic ketoacidosis. N. Engl. J. Med. 300, 11731179.

(20) Yoon, J.W., McClintock, P.R., Onodera, T. and Notkins, A.L. (1980) Virus-induced diabetes mellitus. XVIII. Inhibition by a nondiabetogenic variant of encephalomyocarditis virus. J. Exp. Med. 152, 878-892.

(21) Onodera, T., Taniguchi, T., Yoshihara, K., Shimizu, S., Satoh, M. and Hayashi, T. (1990) Reovirus type 2-induced diabetes in mice prevented by immuno-suppression and effects of thymic hormone. Diabetologia 33, 192-196.

(22) Onodera, T., Yoon, J.W., Brown, K.S. and Notkins, A.L. (1978) Virus-induced diabetes mellitus. XIII. Evidence of genetic control by single locus. Nature 274, 693-696.

(23) Onodera, T., Jenson, A.B., Yoon, J.W. and Notkins, A.L. (1978) Virus-induced diabetes mellitus: Reovirus infection of pancreatic B cells in mice. Science 201, 529-531.

(24) Onodera, T., Toniolo, A., Ray, U.R., Jenson, A.B., Knazek, R.A. and Notkins, A.L. (1981) Virus-induced diabetes mellitus. XX. Polyendocrinopathy and autoimmunity. J. Exp. Med. 153, 14571473.

(25) Onodera, T., Ray, U.R., Meletz, K.A., Suzuki, H., Toniolo, A. and Notkins, A.L. (1982) Autoimmunity and polyendicrine disease prevented by immunosuppression. Nature 297, 66-68.

(26) Come, J.H., Fraser, P.E. and Lansbury, P.T. Jr. (1993) A kinetics model for amyloid formation in the prion diseases: Importance for seeding. Proc. Natl. Acad. Sci. U.S.A. 99, 5959-5963.

(27) Aiken, J.M. and Marsh, R.F. (1990) The search for scrapie agent nucleic acid. Microbiol. Rev. 54, 242-246.

(28) Sakudo, A., Onodera, T., Suganuma, Y., Kobayashi, T., Saeki, K. and Ikuta, K. (2006) Recent advances in clarifying prion protein functions using knockout mice and derived cell lines (Review). Mini Rev. Med. Chem. 6, 589-601.

(29) McKinley, M.P., Taraboulos, A., Kenaga, L., Serban, D., Stieber, A., DeArmond, S.J., Prusiner, S.B. and Gonatas, N. (1991) Ultrastructural localization of kscrapie prion protein in cytoplasmic vesicles of infected cultured cells. Lab. Invest. 65, 622-630.

(30) Jarosz-Griffiths, H.H., Noble, E., Ruthworth, J.V. and Hooper, N.M. (2016) Amyloid-O receptors: The good, the bad, and the prion protein. J. Biol. Chem. 291, 3174-3183.

(31) Pan, K.M., Baldwin, M., Nguyen, J., Gasset, M., Serban, A., Groth, D., Mehlhorn, I., Huang, Z., Fletterick, R.J., Cohen, F.E. and Prusiner, S.B. (1993) Conversion of alpfa-helix into beta-sheets features in the formation of the scrapie prion proteins. Proc. Natl. Acad. Sci. U.S.A. 90, 1096210966.

(32) Mange, A., Beranger, F., Peoc'h, K., Onodera, T., Frobert, Y. and Lehmann, S. (2004) Alpfa- and beta-cleavages of the amino-terminus of the cellular prion protein. Biol. Cell 96, 125-132.

(33) Loudet, D., Dakowski, C., Pietri, M., Pradines, E., Bernard, S., Callebert, J., Aidila-Psorio, H., Mouillet-Richard, S., Launary, J.M., Kellermann, O. and Schneider, B. (2012) Neuritogenesis: the prion protein controls b1 integrin signaling activity. FASEB J. 26, 678-690.

(34) Zhang, C.C., Steele, A.D., Lindquist, S.L. and Lodish, H.F. (2006) Prion protein is expressed on long-term repopulating hematopoietic stem cells and is important for their self-renewal. Proc. Natl. Acad. Sci. U.S.A. 103, 2184-2189.

(35) Steele, A.D., Emsley, J.G., Ozdinler, P.H., Lidquist, S. and Macklis, J. (2006) Prion protein (Pr[P.sup.C]) positively regulates neural precursor proliferation during developmental and adult mammalian neurogenesis. Proc. Natl. Acad. Sci. U.S.A. 103, 3416-3421.

(36) Tremblay, P., Bouzamondo-Bernstein, E., Heinrich, C., Prusiner, S.B. and DeArmond, S.J. (2007) Developmental expression of PrP in the post-implantation embryo. Brain Res. 30, 60-67.

(37) Steele, A.D., Lindquist, S. and Aguzzi, A. (2007) The prion protein knockout mouse: a phenotype under challenge. Prion 1, 83-93.

(38) Kanaani, J., Prusiner, S.B., Diacovo, J., Baekkeskov, S. and Legname, G. (2005) Recombinant prion protein induces rapid polarization and development of synapses in embryonic rat hippocampal neurons in vitro. J. Neurochem. 95, 13731386.

(39) Peralta, O.A., Huckle, W.R. and Eyestona, W.H. (2011) Expression and knockdown of cellular prion protein (Pr[P.sup.C]) in differentiating mouse embryonic stem cells. Differentiation 81, 68--77.

(40) Adle-Biassette, H., Verney, C., Peoc'h, K., Dauge, M.C., Razavi, F., Choudat, L., Gressens, P., Budka, H. and Hein, D. (2006) Immunohistochemical expression prion protein (Pr[P.sup.C]) in the human forebrain during development. J. Neuropathol. Exp. Neurol. 65, 698-706.

(41) Bueler, H., Fischer, M., Lang, Y., Fluethmann, H., Lipp, H.P., DeArmond, S.J., Prusiner, S.B., Aguet, M. and Weissmann, C. (1992) Normal development and behavior of mice lacking the neuronal cell-surface PrP protein. Nature 356, 577-582.

(42) Lopes, M.H., Hajj, G.N., Muras, A.G., Mancini, G.L., Castro, R.M., Ribeiro, K.C., Brentani, R.R., Linden, R. and Martins, V.R. (2005) Interaction of cellular prion and stress-inducible protein 1 promotes neuritogenesis and neuroprotection by distinct signaling pathways. J. Neurosci. 25, 11330-11339.

(43) Zanata, S.M., Lopes, M.H., Mercadante, A.F., Hajj, G.N., Chiarini, L.B., Nomizo, R., Freitas, A.R., Cabral, A.L., Lee, K.S., Juliano, M.A., de Oliveira, E., Jachieri, S.G., Burlingame, A., Huang, L., Linden, R., Brentani, R.R. and Martins, V.R. (2002) Stress-inducible protein 1 is a cell surface ligand for cellular prion that triggers neuroprotection. EMBO J. 21, 3307-3316.

(44) Lassle, M., Blatch, G.L., Kundra, V., Takatori, T. and Zetter, B.R. (1997) Stress-inducible, murine protein mSTI1. Characterization of binding domains for heat shock proteins and in vitro phosphorylation by different kinase. J. Biol. Chem. 272, 1876-1884.

(45) Kramer, M.L., Kratzin, H.D., Schmidt, S., Bomer, A., Windl, O., Liemann, S., Hornemann, S. and Kretzschmar, H. (2001) Prion protein binds copper within the physiological concentration range. J. Biol. Chem. 276, 16711-16719.

(46) Sakudo, A., Lee, D., Yoshimura, E., Nagasaka, S., Nitta, K., Saeki, K., Matsumoto, Y., Lehmann, S., Itohara, S., Sakaguchi, S. and Onodera, T. (2004) Prion protein suppress perturbation of cellular copper homeostasis under oxidative conditions. Biochem. Biophys. Res. Commun. 313, 850-855.

(47) Prusiner, S.B. (1997) Prion diseases and BSE crisis. Science 278, 245-251.

(48) Brown, D.R., Qin, K., Herms, J.W., Madlung, A., Manson, J., Strome, R., Fraser, P.E., Kruck, T., Von Bohlen, A., Schulz-Schaeffer, W., Giese, A., Westaway, D. and Kretzschmar, H. (1997) The cellular prion protein binds copper in vitro. Nature 390, 684-687.

(49) Kretzschmar, H.A., Tings, T., Madlung, A., Giese, A. and Herms, J. (2000) Function of PrP(C) as a copper-binding protein at the synapse. Arch. Virol. Suppl. 16, 239-249.

(50) Brown, D.R. (1999) Prion protein expression aids cellular uptake and veratridine-induced release of copper. J. Neurosci. Res. 58, 717-725.

(51) Haigh, C.L., Edwards, K. and Brown, D.R. (2005) Copper binding is governing determinant of prion protein turnover. Mol. Cell. Neurosci. 30, 186196.

(52) Kubosaki, A., Nishimura-Nasu, Y., Nishimura, T., Yusa, S., Sakudo, A., Saeki, K., Matsumoto, Y., Itohara, S. and Onodera, T. (2003) Expression of normal cellular prion protein (Pr[P.sup.C]) on T lymphocytes and effect of copper ion: analysis by wild-type and prion gene-deficient mice. Biochem. Biophys. Res. Commun. 307, 810-813.

53) Kuwahara, C., Takeuchi, A.M., Nishimura, T., Haraguchi, K., Kubosaki, A., Matsumoto, Y., Saeki, K., Matsumoto, Y., Yokoyama, T., Itohara, S. and Onodera, T. (1999) Prions prevent neuronal cell-line death. Nature 400, 225-226.

(54) Sakudo, A., Lee, D.C., Saeki, K., Nakamura, Y., Inoue, K., Matsumoto, Y., Itohara, S. and Onodera, T. (2003) Impairment superoxide dismutase activation by N-terminally truncated prion protein (PrP) in PrP-deficient neuronal cell line. Biochem. Biophys. Res. Commun. 308, 660-667.

(55) Sakudo, A., Nakamura, I., Ikita, K. and Onodera, T. (2007) Recent developments in prion disease research: diagnostic tools and in vitro cell culture models. J. Vet. Med. Sci. 69, 329-337.

(56) Sakudo, A., Onodera, T. and Ikuta, K. (2007) Prion protein gene deficient cell lines: powerful tools for prion biology. Microbiol. Immunol. 51, 1-13.

(57) Nishimura, T., Sakudo, A., Hashiyama, Y., Yachi, A., Saeki, K., Matsumoto, Y., Ogawa, M., Sakaguchi, S., Itohara, S. and Onodera, T. (2007) Serum withdrawal-induced apoptosis in ZrchI prion protein (PrP) gene-deficient neuronal cell line is suppressed by PrP, independent of Doppel. Microbiol. Immunol. 51, 457-466.

(58) Pattison, I.H. and Jebbett, J.N. (1973) Clinical and histological recovery from the scrapie-like spongiform encephalopathy produced in mice by feeding them with cuprizone. J. Pathol. 109, 245-250.

(59) Bounhar, Y., Zhang, Y., Goodyer, C.G. and LeBlanc, A. (2001) Prion protein protects human neurons against Bax-mediated apoptosis. J. Biol. Chem. 276, 39145-39149.

(60) Roucou, X. and LeBlanc, A.C. (2005) Cellular prion protein neuroprotective function: implication in prion diseases. J. Mol. Med. 83, 3-11.

(61) Schneider, B., Mutel, V., Pietri, M., Ermonval, M., Mouillet-Richard, S. and Kellermann, O. (2003) NADPH oxidase are extracellular regulated kinase 1/2 are targets of prion protein signaling in neuronal and nonneuronal cells. Proc. Natl. Acad. Sci. U.S.A. 100, 13326-13331.

(62) Spielhaupter, C. and Schatzl, H.M. (2001) Pr[P.sup.C] directly interacts with protein involved in signaling pathways. J. Biol. Chem. 276, 44604-44612.

(63) Meggio, F., Negro, A., Sarno, S., Ruzzene, M., Bertoli, A., Sorgato, M.C. and Pinna, L.A. (2000) Bovine prion protein as a modulator of protein kinase CK2. Biochem. J. 352, 191-196.

(64) Chiarini, L.B., Freitas, A.R.O., Zanata, S.M., Brentani, R.R., Martins, V.R. and Linden, R. (2002) Cellular prion protein transduces neuroprotective signals. EMBO J. 21, 3317--3326.

(65) Vassallo, N., Herms, J., Behrens, C., Krebs, B., Saeki, K., Onodera, T., Windl, O. and Kretzschmar, H. (2005) Activation of phospatidylinositol 3-kinase by cellular prion protein and its role in cell survival. Biochem. Biophys. Res. Commun. 332, 75-82.

(66) Hoshino, S., Inoue, K., Yokoyama, T., Kobayashi, Asakura, T., Teramoto, A. and Itohara, S. (2003) Prions prevent brain damage after experimental brain injury: a preliminary report. Acta Neurochir. Suppl. 86, 297-299.

(67) Shyu, W.C., Chen, C.P., Saeki, K., Kubosaki, A., Matsumoto, Y., Onodera, T., Ding, D.C., Chiang, M.F., Lee, Y.J., Lin, S.Z. and Li, H. (2005) Hypoglycemia enhances the expression of prion protein and heat-shock protein 70 in a mouse neuroblastoma cell line. J. Neurosci. Res. 80, 887894.

(68) Sakudo, A., Lee, D.C., Nishimura, T., Li, S., Tsuji, Nakamura, T., Matsumoto, Y., Saeki, K., Itohara, S., Ikuta, K. and Onodera, T. (2005) Octapeptide repeat region and N-terminal half of hydrophobic region of prion protein (PrP) mediates PrP-dependent activation of superoxide dismutase. Biochem. Biophys. Res. Commun. 326, 600-606.

(69) Hashimoto, A., Onodera, T., Ikeda, H. and Kitani, H. (2000) Isolation and characterization of fetal bovine brain cells in primary culture. Res. Vet. Sci. 69, 39-46.

(70) Moore, R.C., Lee, I.Y., Silverman, G.L., Harrison, P.M., Strome, R., Heinrich, C., Karunaratne, A., Pasternak, S.H., Chishti, M.A., Liang, Y., Mastrangelo, P., Wang, K., Smit, A.F., Katamine, S., Carlson, G.A., Cohen, F.E., Prusiner, S.B., Melton, D.W., Tremblay, P., Hood, L.E. and Westaway, D. (1999) Ataxia in prion protein (PrP)-deficient mice is associated with upregulation of the novel PrP-like protein doppel. J. Mol. Biol. 292, 797-817.

(71) Rossi, D., Cozzio, A., Flechsig, E., Klein, M.A., Rulicke, T., Aguzzi, A. and Weissmann, C. (2001) Onset of ataxia and Purkinje cell loss in PrP null mice inversely correlated with Dpl level in brain. EMBO J. 20, 694-702.

(72) Sakaguchi, S., Katamine, S., Nishida, N., Moriuchi, R., Shigematsu, K., Sugimoto, T., Nakatani, A., KatA[beta]ka, Y., Houtani, T., Shirabe, S., Okada, H., Hasegawa, S., Miyamoto, T. and Noda, T. (1996) Loss of cerebellar Purkinje cells in aged mice homozygous for a disrupted PrP gene. Nature 380, 528-531.

(73) Ersdal, C., Ulvund, M.J., Benestad, S.L. and Tianulis, M.A. (2003) Accumulation of pathogenic prion protein (Pr[P.sup.Sc]) in nervous and lymphoid tissue of sheep with subclinical scrapie. Vet. Pathol. 40, 164-174.

(74) Park, J.S., Onodera, T., Nishimura, S., Thompson, R.F. and Itohara, S. (2006) Molecular evidence for two-stage learning and partial laterality in eyeblink conditioning of mice. Proc. Natl. Acad. Sci. U.S.A. 103, 5549-5554.

(75) Solforosi, L., Criado, J.R., McGavern, D.B., Wirz, S., Sanchez-Alavez, M., Sugama, S., DeGiorgio, L.A., Volpe, B.T., Wiseman, E., Abalos, G., Masliah, E., Gilden, D., Oldstone, M.B., Conti, B. and Williamson, R.A. (2004) Cross-linking cellular prion protein triggers neuronal apoptosis in vivo. Science 303, 1514-1516.

(76) Winklhofer, K.F., Tatzelt, J. and Haass, C. (2008) The two faces of protein misfolding: gain-and lossof-function in neurodegenerative diseases. EMBO J. 23, 336-349.

(77) Bueler, H., Aguzzi, A., Sailer, A., Greiner, R.A., Autenried, P., Aguet, M. and Weissmann, C. (1993) Mice devoid of PrP are resistant to scrapie. Cell 73, 1339-1347.

(78) Kim, C.K., Sakudo, A., Taniuchi, Y., Shigematsu, K., Kang, C.B., Saeki, K., Matsumoto, Y., Sakaguchi, S., Itohara, S. and Onodera, T. (2007) Late-onset olfactory deficits and mitral cell loss in mice lacking prion protein with ectopic expression of Doppel. Int. J. Mol. Med. 20, 169176.

(79) Le Pichon, C.E., Valley, M.T., Polymenidou, M., Chesler, A.T., Sagdullaev, B.T., Aguzzi, A. and Firestein, S. (2009) Olfactory behavior and physiology are disrupted in prion protein knockout mice. Nat. Neurosci. 12, 60-69.

(80) Manson, J.C., Clarke, A.R., Hooper, M.L., Aitchinson, L., McConnell, I. and Hope, J. (1994) 129/Ola mice carrying a null mutation in PrP that abolishes mRNA productions are developmentally normal. Mol. Neurobiol. 8, 121-127.

(81) Lledo, P.M. and Lagier, S. (2006) Adjusting neurophysiological computation in the adult olfactory bulb. Semin. Cell Dev. Biol. 17, 443-453.

(82) Stopfer, M. (2007) Olfactory processing: massive convergence onto sparse codes. Curr. Biol. 17, R363-R364.

(83) Kashiwadani, H., Sasaki, Y.F., Uchida, N. and Mori, K. (1999) Synchronized oscillatory discharge of mitral/tufted cells with different molecular receptive ranges in the rabbit olfactory bulb. J. Neurophysiol. 82, 1786-1792.

(84) Beshel, J., Kopell, N. and Kay, L.M. (2007) Olfactory bulb gamma oscillations are enhanced with task demands. J. Neurosci. 27, 8358-8365.

(85) Urban, N.N. (2002) Lateral inhibition in the olfactory bulb and in olfaction. Physiol. Behav. 77, 607-612.

(86) Yokoi, M., Mori, K. and Nakanishi, S. (1995) Refreshment of odor molecule tuning by dendrodendritic synaptic inhibition in the olfactory bulb. Proc. Natl. Acad. Sci. U.S.A. 92, 3371-3375.

(87) Shepherd, G.M. (2003) The Synaptic Organization of Brain. Oxford University Press, New York, U.S.A.

(88) Lagier, S., Panzanelli, P., Russo, R.E., Nissant, A., Bathellier, B., Sassoe-Pongnetto, M., Fritschy, J.M. and Lledo, P.M. (2007) GABAergic inhibition at dendrodendritic synapse tunes gamma oscillations in the olfactory bulb. Proc. Natl. Acad. Sci. U.S.A. 104, 7259-7264.

(89) Linden, R., Martins, V.R., Prado, M.A., Cammarota, M., Izquierdo, I. and Brentani, R.R. (2008) Physiology of the prion protein. Physiol. Rev. 88, 673-728.

(90) Prestori, F., Rossi, P., Bearzatto, B., Laine, J., Necchi, D., Diwaker, S., Schiffmann, S.N., Axelrad, H. and D'Angelo, E. (2008) Altered neuron excitability and synaptic plasticity in the cerebellar granular layer of juvenile prion protein knock-out mice with impaired motor control. J. Neurosci. 28, 7091-7103.

(91) Nishida, N., Tremblay, P., Sugimoto, T., Shigematsu, K., Shirabe, S., Petromilli, C., Erpel, S.P., Nakaoke, R., Atarashi, R., Houtani, T., Torchia, M., Sakaguchi, S., DeArmond, S.J., Prusiner, S.B. and Katamine, S. (1999) A mouse prion protein transgene rescues mice deficient for the prion protein gene from Purkinje cell degeneration and demyelination. Lab. Invest. 79, 689-697.

(92) Bremer, J., Baumann, F., Tiberi, C., Wessig, C., Fischer, H., Schwarz, P., Steele, A.D., Toyka, K.V., Nave, K.-A., Weis, J. and Aguzzi, A. (2010) Axonal prion protein is required for peripheral myelin maintenance. Nat. Neurosci. 13, 310-318.

(93) Heikenwalder, M., Kurrer, M.O., Margalith, I., Kranich, J., Zeller, N., Haybaeck, J., Polymenidou, M., Matter, M., Bremer, J., Jackson, W.S., Lindquist, S., Sigurdson, C.J. and Aguzzi, A. (2008) Lymphotoxin-dependent prion replication in inflammatory stromal cells of granulomas. Immunity 29, 998-1008.

(94) Genoud, N., Behrens, A., Miele, G., Robay, D., Heppner, F.L., Freigang, S. and Aguzzi, A. (2004) Disruption of Doppel prevents neurodegeneration in mice with extensive Prnp deletions. Proc. Natl. Acad. Sci. U.S.A. 101, 4198-4203.

(95) Behrens, A., Genoud, N., Naumann, H., Rulicke, T., Janett, F., Heppner, F.L., Ledermann, B. and Aguzzi, A. (2002) Absence of the prion protein homologue Doppel causes male sterility. EMBO J. 21, 3652-3658.

(96) Rutishauser, D., Mertz, K.D., Moos, R., Brunner, E., Rulicke, T., Calella, A.M. and Aguzzi, A. (2009) The comprehensive native interactome of a fully functional tagged prion protein. PLoS One 4, e4446.

(97) Baumann, F., Tolnay, M., Brabeck, C., Pahnke, J., Kloz, U., Niemann, H.H., Heikenwalder, M., Rulicke, T., Burkle, A. and Aguzzi, A. (2007) Lethal recessive myelin toxicity of prion protein lacking its central domain. EmBo J. 26, 538-547.

(98) Watt, N.T. and Hooper, N.M. (2005) Reactive oxygen species (ROS)-mediated beta-cleavage of the prion protein in the mechanism of the cellular response to oxidative stress. Biochem. Soc. Trans. 33, 1123-1125.

(99) Sunyach, C., Cisse, M.A., da Costa, C.A., Vincent, B. and Checler, F. (2007) The C-terminal products of cellular prion protein processing, C1 and C2, exert distinct influence on p53-dependent staurosporine-induced caspase-3 activation. J. Biol. Chem. 282, 1956-1963.

(100) Walmsley, A.R., Watt, N.T., Taylor, D.R., Perera, W.S. and Hooper, N.M. (2009) Alpha-cleavage of the prion protein occurs in a late compartment of the secretory pathway and is independent of lipid rafts. Mol. Cell. Neurosci. 40, 242-248.

(101) Chesebro, B., Trifilo, M., Race, R., Meadr-White, K., Teng, C., LaCasse, R., Raymond, L., Favara, C., Baron, G., Priola, S., Caughey, B., Masliah, E. and Oldstone, M. (2005) Anchorless prion protein results in infectious amyloid disease without clinical scrapie. Science 308, 1435-1439.

(102) Nazor, K.E., Seward, T. and Telling, G.C. (2007) Motor behavioral and neuropathological deficits in mice for normal prion protein expression. Biochim. Biophys. Acta 1772, 645-653.

(103) Radvanivic, I., Braun, N., Giger, O.T., Mertz, K., Miele, G., Prinz, M., Navarro, B. and Aguzzi, A. (2005) Truncated prion protein and Doppel are myelinotoxic in the absence of oligodendrocytic Pr[P.sup.C]. J. Neurosci. 25, 4879-4888.

(104) Collinge, J., Whittington, M.A., Sidle, K.C., Smith, C. J., Palmer, M.S., Clarke, A.R. and Jefferys, J.G. (1994) Prion protein is necessary for normal synaptic function. Nature 370, 295-297.

(105) Mallucci, G.R., Ratte, S., Asante, E.A., Linehan, J., Gowland, I., Jefferys, J.G. and Collinge, J. (2002) Postnatal knock out of prion protein alters hippocampal CA1 properties, but not result in neurodegeneration. EMBO J. 21, 202-210.

(106) Issacs, J.D., Jackson, G.S. and Altmann, D.M. (2006) The role of the cellular prion protein in the immune system. Clin. Exp. Immunol. 146, 1-8.

(107) Kuffer, A., Lakkaraju, A.K.K., Mogha, A., Petersen, S.C., Airich, K., Doucerain, C., Marpakwar, R., Bakirci, P., Senatore, A., Monnard, A., Schiav, C., Nuvolone, M., Grosshans, B., Hornemann, S., Bassilana, F., Monk, K.R. and Aguzzi, A. (2016) The prion protein is an agonistic ligand of the G protein-coupled receptor Adgrg6. Nature 536, 464-468.

(108) Safar, J.G., Geschwind, M.D., Deering, C., Didorenko, S., Sattavat, M., Sanchez, H., Servan, A., Vey, M., Baron, H., Giles, K., Miller, B.L., DeArmond, S.J. and Prusiner, S.B. (2005) Diagnosis of human prion disease. Proc. Natl. Acad. Sci. U.S.A. 102, 3501-3506.

(109) White, M.D., Farmer, M., Mirabile, I., Brander, S., Collinge, J. and Mallucci, G.R. (2008) Sigle treatment with RNAi against prion protein rescues early neuronal dysfunction and prolongs survival in mice with prion disease. Proc. Natl. Acad. Sci. U.S.A. 105, 10238-10243.

(110) Moreno, J.A., Radford, H., Peretti, D., Steinert, J.R., Verity, N., Martin, M.G., Halliday, M., Morgan, J., Dinadale, D., Ortori, C.A., Barret, D. A., Tsaytler, P., Bertolotti, A., Willis, A.E., Bushell, M. and Mallucci, G.R. (2012) Sustained translational repression by eIF2a-P mediates prion neurodegeneration. Nature 485, 507-511.

(111) Tsayler, P., Harding, H.P., Ron, D. and Bertolotti, A. (2011) Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis. Science 332, 91-94.

(112) Balch, W.E., Morimoto, R.I., Dillin, A. and Kelly, J.W. (2008) Adapting proteostasis for disease intervention. Science 319, 916-919.

(113) Ma, J., Wollmann, R. and Lindquist, S. (2002) Neurotoxicity and neurodegeneration when PrP accumulates in the cytosol. Science 298, 1781-1785.

(114) Wang, X., Wang, F., Arterburn, L., Wollmann, R. and Ma, J. (2006) The interaction between cytoplasmic prion protein and the hydrophobic lipid core of membrane correlates with neurotoxicity. J. Biol. Chem. 281, 13559-13565.

(115) Rambold, A.S., Miesbauer, M., Rapaport, D., Bartke, T., Baier, M., Winklhofer, K.F. and Tatzelt, J. (2006) Association of Bcl-2 with misfolded prion protein is linked to the toxic potential of cytosolic PrP. Mol. Biol. Cell 17, 3356-3368.

(116) Kristiansen, M., Deriziotis, P., Dimcheff, D.E., Jackson, G.S., Ovaa, H., Naumann, H., Clarke, A.R., Van Leeuwen, F.W., Menendez-Benito, V., Dantuma, N.P., Portis, J.L., Collinge, J. and Tabrizi, S.J. (2007) Disease-associated prion oligomers inhibit the 26S proteasome. Mol. Cell 26, 175-188.

(117) Mok, S.W., Riemer, C., Madela, K., Hsu, D.K., Liu, F.T., Gultner, S., Heise, I. and Baier, M. (2007) Role of galectin-3 in prion infections of the CNS. Biochem. Biophys. Res. Commun. 359, 672-678.

(118) Guillot-Sestier, M.V. and Cheeler, F. (2012) Cellular prion and its catabolites in the brain: production and function. Curr. Mol. Med. 12, 304-315.

(119) Onodera, T., Sakudo, A., Tsubone, H. and Itohara, S. (2014) Review of studies that have used knockout mice to assess normal function of prion protein under immunological or pathophysiological stress. Microbiol. Immunol. 58, 361-374.

(120) Cirado, J.R., Sanchez-Alavez, M., Conti, B., Giacchino, J.L., Wills, D.N., Henriksen, S.J., Race, R., Manson, J.C., Chesebro, B. and Oldstone, M.B. (2005) Mice devoid of prion protein have cognitive deficits that are rescued by reconstitution of PrP in neurons. Neurobiol. Dis. 19, 255-265.

(121) Weise, J., Sandau, R., Schwarting, S., Crome, B., Wrede, A., Schultz-Schaeffer, W., Zerr, I. and Bahr, M. (2006) Deletion of cellular prion protein results in Akt activation, enhanced postischemic caspase-3 activation, and exacerbation of ischemic brain injury. Stroke 37, 1296-1300.

(122) Guillot-Sestier, M.V., Sunyach, C., Druon, C., Scarzello, S. and Checler, F. (2009) The alpha-secretase-derived N-terminal product of cellular prion, N1, displays neuroprotective function in vitro and in vivo. J. Biol. Chem. 284, 35973-35986.

(123) Westaway, D., DeArmond, S.J., Cayetano-Calas, J., Groth, D., Foster, D., Yang, S.L., Torchia, M., Carlson, G.A. and Prusiner, S.B. (1994) Degeneration of skeletal muscle, peripheral nerves, and central nervous system in transgenic mice over-expressing wild-type prion proteins. Cell 76, 117-129.

(124) Brandner, S., Isenmann, S., Raeber, A., Fischer, M., Sailer, A., Kobayashi, Y., Marino, S. and Weissmann, C. (1996) Normal host prion protein is necessary for scrapie-induced neurotoxicity. Nature 379, 339-343.

(125) Lauren, J., Gimble, D.A., Nygaard, H.B., Gilbert, J. W. and Strittmatter, S.M. (2009) Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers. Nature 457, 1128-1132.

(126) Gimbel, D.A., Nygaard, H.B., Coffey, E.E., Gunther, E.G., Lauren, J., Gimbel, Z.A. and Strittmatter, S.M. (2010) Memory impairment in transgenic Alzheimer mice require cellular prion protein. J. Neurosci. 30, 6367-6374.

(127) Balducci, C., Beeg, M., Stravalaci, M., Bastone, A., Sclip, A., Biasini, E., Tapella, L., Colombo, L., Manzoni, C., Borsello, T., Chiesa, R., Gobbi, M., Salmonea, M. and Forloni, G. (2010) Synthetic amyloid-beta oligomers impair long-term memory independently of cellular prion protein. Proc. Natl. Acad. Sci. U.S.A. 107, 2295-2300.

(128) Stys, P.K., You, H. and Zamponi, G.W. (2012) Copper-dependent regulation of NMDA receptors by cellular prion protein: implication for neuro-degenerative disorders. J. Physiol. 590, 1357-1368.

(129) Parkin, E.T., Watt, N.T., Hussain, I., Eckman, E.A., Eckman, C.B., Manson, J.C., Baybutt, H.N., Turner, A.J. and Hooper, N.M. (2007) Cellular prion protein regulates beta-secretase cleavage of the Alzheimer's amyloid precursor protein. Proc. Natl. Acad. Sci. U.S.A. 104, 11062-11067.

(130) Cisse, M.A., Sunyach, C., Lefranc-Jullien, S., Postina, R., Vincent, B. and Checler, F. (2005) The disintegrin ADAM9 indirectly contributes to the physiological processing of cellular prion by modulating ADAM10 activity. J. Biol. Chem. 280, 40624-40631.

(131) Schmitt-Ulms, G., Legname, G., Baldwin, M.A., Ball, H.L., Bradon, N., Bosque, P.J., Crossin, K. L., Edelman, G.M., DeArmond, S.J., Cohen, F.E. and Prusiner, S.B. (2001) Binding of neural cell adhesion molecules (N-CAMs) to the cellular prion protein. J. Mol. Biol. 314, 1209-1225.

(132) Santuccione, A., Sytnyk, V., Leshchyns'ka, I. and Schachner, M. (2005) Prion protein recruits its neuronal receptor NCAM to lipid rafts to activate p59fyn and to enhance neurite outgrowth. J. Cell Biol. 169, 341-354.

(133) Taylor, D.R. and Hooper, N.M. (2007) The low density lipoprotein receptor-related protein 1 (LRP1) mediates the endocytosis of the cellular prion protein. Biochem. J. 402, 17-23.

(134) Parkyn, C.J., Vermeulen, E.G., Mootoosamy, R.C., Sunyach, C., Lacobsen, C., Oxvig, C., Moestrup, Liu, Q., Bu, G., Jen, A. and Morris, R.J. (2008) LRP1 control biosynthetic and endocytic trafficking of neural prion protein. J. Cell Sci. 121, 773783.

(135) Hajj, G.N., Lopes, M.H., Mercadante, A.F., Veiga, da Silveira, R.B., Santos, T.G., Ribeiro, K.C., Juriano, M.A., Jacchieri, S.G., Zanata, S.M. and Martins, V.R. (2007) Cellular prion protein interaction with vitronectin supports axonal growth and is compensated by integrins. J. Cell Sci. 120, 1915-1926.

(136) Sakudo, A., Lee, D.C., Li, S., Nakamura, T., Matsumoto, Y., Saeki, K., Itohara, S., Ikuta, K. and Onodera, T. (2005) PrP cooperates with STI1 to regulate SOD activity in PrP deficient neuronal cell line. Biochem. Biophys. Res. Commun. 328, 14-19.

(137) Martins, V.R., Beraldo, F.H., Hajj, G.N., Lopes, M.H., Lee, S.K., Prado, M.M. and Linden, R. (2010) Prion protein: orchestrating neurotrophic activities. Curr. Issues Mol. Biol. 12, 63-86.

(138) Nah, J., Pyo, J.O., Jung, S., Yoo, S.M., Kam, T.I., Chang, J.W., Han, J., An, S.S.A., Onodera, T. and Jung, Y.K. (2013) BECN1/Beclin is recruited into lipid rafts by prion to activate autophagy in response to amyloid A[[BETA].SUB.42]. Autophagy 9, 2009-2022.

(139) Keshet, G.I., Ovaida, H., Taraboulos, A. and Gabison, R. (1999) Scrapie-infected mice and PrP knockout mice share abnormal localization and activity of neuronal nitric oxide synthase. J. Neurochem. 72, 1224-1231.

(140) Nasu-Nishimura, Y., Taniuchi, Y., Nishimura, T., Sakudo, A., Nakajima, K., Ano, Y., Sugiura, K., Sakaguchi, S., Itohara, S. and Onodera, T. (2008) Cellular prion protein prevents brain damage after encephalomyocarditis virus (EMCV) infection in mice. Arch. Virol. 153, 1007-1012.

(141) Kitamura, Y., Yanagisawa, D., Inden, M., Tanaka, K., Tsuchiya, D., Kawasaki, T., Taniguchi, T. and Shimohama, S. (2005) Recovery of focal brain ischemia-induced behavior dysfunction by intracerebroventricular injection of microglia. J. Pharmacol. Sci. 97, 289-293.

(142) Dal-Pizzol, F., Klamt, F., Vianna, M.M., Schoroder, N., Quevedo, J., Benfato, M.S., Moreira, J.C. and Waltz, R. (2000) Lipid peroxidation in hippocampus early and late after status epilepticus induced by pilocarpine or kainic acid in Wistar rats. Neurosci. Lett. 291, 179-182.

(143) Hafiz, F.B. and Brown, D.R. (2000) A model for the mechanism of astroglia in prion diseases. Mol. Cell. Neurosci. 16, 221-232.

(144) Wong, B.S., Liu, T., Li, R., Pan, T., Petersen, R.B., Smith, M.A., Gambetti, P., Perry, G., Manson, J.C., Brown, D.R. and Sy, M.S. (2001) Increased levels of oxidative stress markers detected in the brain of mice devoid of prion proteins. J. Neurochem. 76, 565-572.

(145) Sriskandan, S. and Altmann, D.M. (2008) The immunology of sepsis. J. Pathol. 214, 214-223.

(146) De Almeida, C.J., Chiarini, L.B., da Silva, J.P., Silva, E.P.M., Marius, M.A. and Linden, R. (2005) The cellular prion protein modulates phagocytosis and inflammatory response. J. Leukoc. Biol. 77, 238-246.

(147) Zhou, R., Tardivel, A., Thorens, B., Choi, I. and Tschopp, J. (2010) Thioredoxin-interacting protein links oxidative stress in inflasome activation. Nat. Immunol. 11, 136-140.

(148) Brown, P., Will, R.G., Bradley, R., Asher, D.M. and Detwiler, L. (2001) Bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease: background, evolution and current concerns. Emerg. Infect. Dis. 7, 6-16.

(149) Sakudo, A. and Onodera, T. (2015) PrP gene knockout cell line: insight into functions of the PrP. Front. Cell Dev. Biol. 15, 2-75, doi: 10.3389/fcell.2014.00075.

(Received Nov. 2, 2016; accepted Jan. 26, 2017)


Takashi Onodera was born in 1947. He graduated from the University of Tokyo, School of Agricultural and Life Sciences in 1969 with a doctorate degree in veterinary medicine (DVM). After his graduation from the veterinary school, he majored in zoonosis and received his Ph.D. degree in the Graduate Student Training Program in 1974 from the Institute of Medical Science, and Graduate School of Agricultural and Life Sciences of the University of Tokyo. He worked as an Assistant Professor at the Department of Animal Pathology, Institute of Medical Science, in the University of Tokyo between 1974-1977. He was given a Visiting Fellowship at the National Institute of Dental Research (NIDR), National Institutes of Health (NIH) in the United States (U.S.A.) in 1974. In 1977 he was a Visiting Associate in the Laboratory of Oral Medicine (LOM) in NIDR, NIH, and in 1981 he was promoted to serve as an Expert in LOM, NIDR, NIH. During his days at NIH, U.S.A., he worked for Dr. Abner Louis Notkins, Chief of LOM, to investigate the virus-induced diabetes mellitus and autoimmunity. Upon his accomplishment, he received the NIH Director's Award (titled: Innovative studies on the role of viruses in autoimmunity) in 1983. He was appointed as the Chief of Laboratory of Immunology, National Institute of Animal Health in Ministry of Agriculture, Forestry and Fisheries (MAFF) in Japan in 1986, and served as the Chief of Laboratory of Immune Cytology, National Institute of Animal Health in 1988. He assumed a chair, and became the Professor and Chairman of the Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo in Tokyo, Japan (1991-2010). During his career in MAFF between 1984-1991, he received the Bifidus Award from the Japanese Society for Intestinal Microbiology for his study of prion diseases. He has a distinguished research career in unfurling the virological and immunopathological mechanisms controlling slow virus infection as well as contributing to the development of prion infectiology. At the U.S. NIH, his pioneering works covered a broad range of immunopathology of virus infections, such as in the fields of dentistry, virology and vaccinology. While in Japan, his scientific contribution has focused on the field of slow virus associated with normal function of prion protein in prion diseases and Alzheimer's diseases, and epidemiological analysis of Japanese BSE. Currently he is working on chemicals to treat and prevent for prions and Alzheimer's disease. Besides his career at the University of Tokyo, he served as a Government Advisor for MAFF, Ministry of Health, Labor and Welfare, and Cabinet Office. He assumed the Chair in the Advisory Committee on Food, Agriculture and Rural Policy's Subcommittee on Prion Disease, MAFF in 2000, and became a Member of the Expert Committee on Prions, Food Safety Commission in the Cabinet Office in 2003. During his career as a Chairperson in MAFF, the Prion Disease Subcommittee spotted the first Japanese BSE case, which was also the first domestic BSE case outside Europe. For these achievements and scientific contributions, he was awarded the Japan Agricultural Science Award, and the Yomiuri Agriculture Award (titled: Animal model for prion diseases) in 2009. He is now a Project Professor in Research Center for Food Safety, University of Tokyo (2010--present), and is also working as a Visiting Scientist in the Brain Research Institute of Riken (1994--present). He has served as a Member in Institut de France (French National Academy of Science and Art, titled: Veterinary Science) in Paris since 2012, and is still actively duty-bound to said position. He further serves as a Board Member of the Japanese Society of Bio-defense and of the Human Genome Research Center, Institute of Medical Science of the University of Tokyo. He has also served as a President for the following organizations: the Japanese Society for Neurovirology, and the Japanese Society for Veterinary Immunology.

Photographer: Mayumi Suzuki

By Takashi ONODERA * [1], ([dagger])

(Communicated by Masanori OTSUKA, M.J.A.)

* [1] Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, Japan.

([dagger]) Correspondence should be addressed: T. Onodera, Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, the University of Tokyo, 1-1-1 Yayoi, Bunkyoku, Tokyo 113-8657, Japan (e-mail:

Abbreviations: A[beta]: [beta]-amyloid; A[beta]O: A[BETA] oligomer; [alpha]7nAChR: [alpha]7 nicotinic acetylcholine receptor; AD: Alzheimer's disease; Bcl-2: B-cell lymphoma 2 protein; BECN: beclin; BSE: bovine spongiform encephalopathy; cAMP: cyclic adenosine monophosphate; CDP: chronic demyelinating polyneuropathy; CJD: Creutzfeldt-Jakob disease; CK2: protein kinase CK2; CWD: chronic wasting disease; cytoPrP: cytosolic compartment of prion protein; DCN: deep cerebellar nuclei; Dpl: doppel; EMCV: encephalomyocarditis virus; ERK: extracellular signal-regulated kinase; GPI: glycophosphatidylinositol; Grb2: growth factor receptor bound protein 2; HIV: human immunodeficiency virus; Hsp: heat shock protein; HTLV: human T-cell lymphotropic virus; IPSP: inhibitory postsynaptic potential; LFP: local field potential; LPR1: low-density lipoprotein-related protein 1; LTP: long-term potentiation; MAPK: mitogen-activated protein kinase; mGluR5: metabotropic glutamate receptor 5; NADPH: nicotinamide adenine dinucleotide phosphate; NLRP3: nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing3; NMDAR: N-methyl-D-asparate receptor; NO: nitric oxide; NOS: nitric oxide synthase; OB: olfactory bulb; PI3K: phosphatidylinositol 3-kinase; PI3KC3: phosphatidylinositol 3-kinase, catalytic subunit type 3; PKA: protein kinase A; PML: progressive multifocal leukoencephalopathy; Pint1: prion protein interacting protein 1; Prnp: prion protein gene; PRNP: human prion protein gene; PrP: prion protein; Pr[P.sup.C]: cellular isoform of prion protein; Pr[P.sup.Sc]: scrapie isoform of prion protein; Rag1: recombination activating gene 1; ROS: reactive oxygen species; RSSE: Russian spring summer encephalitis; SOD: super oxide dismutase; STI1: stress inducible protein 1; TCA: transmissible cerebral amyloidosis; []GPI-PrP: GPI deficient PrP transgenic mouse; TSE: transmissible spongiform encephalopathy; UPR: unfolded protein response; UPS: ubiquitin-proteasome system.

Caption: Fig. 1. Number of BSE cases in the United Kingdom (U.K.) and Japan. An estimated 180,000 cattle were affected by BSE and killed in U.K. As a result, 156 people died in the 1990s as after having contracted the variant of CJD. To date, that number of human mortalities has risen to 177. In 1992, 37280 BSE cases were observed. Due to introduction of Feed ban for Specified Bovine Offal in 1988, number of BSE cases decreased from 1993 in [U.K..sup.148]) In Japan, 36 cattle found to be affected by BSE, and were subsequently sacrificed between 2001--2009.

Caption: Fig. 2. Neuroprotective function of cellular prion protein. Laminin receptor precursor (LRP) and Stress-inducible protein 1 (STI1) are the important signals protecting neurons from apoptosis. STI1 protective signals were mediated by cAMP/PKA pathway or MAPK/ ERK pathway. Oncogene products Src also mediates protective signals through PI3K.

Caption: Fig. 3. Phenotypes of [Prnp.sup.-/-] mice established in several laboratories in the world. Different kinds of targeting cassette for [Prnp.sup.-/-] are shown. The structures of constructs used to produce six lines of [Prnp.sup.-/-] mice are explained. The structure of wild-type (WT) [Prnp.sup.-/-] exon3 and PrP coding region (open box) is shown at the top. The selection markers are indicated by black boxes. The presence and absence of 3 splicing acceptors are correlated with ectopic expression of Doppel (Dpl) and development of late-onset ataxia induced by loss of Purkinje cells. Chronic demyelinating polyneuropathy (CDP) was observed in type-1 and type-2 [Prnp.sup.-/-] mice. The selection markers were: PGK, mouse phosphoglycerate kinase promoter; NEO, neomycin phosphotransferase; HPRT, mouse hypoxanthine phosphoribosyltransferase; TK, human herpes simplex virus type 1 thymidine kinase promoter; MT, mouse metallothionein promoter; loxP, a 34 bp recombination site from phage P1. The restriction enzyme sites were: E, EcoRI; X, Xbal, S.A., splicing acceptor.149) Zurich type-2 knockout mouse produces while type-1 mouse does not produce doppel proteins. NT: not tested.

Caption: Fig. 4. Capacity of truncated [Prnp.sup.-/-] genes to rescue apoptosis when reintroduced to [Prnp.sup.-/-] cells. Introducing empty vector (EM) to [Prnp.sup.-/-] cells shows strong apoptosis. However, introducing [DELTA]#1 (octa-repeat region (OR): aa 53-94 truncated) induces more potent apoptosis, showing the dominant negative effect of [DELTA]#1 gene. Similarly, introducing [DELTA]#2 (STI1 binding area; N-terminal half of hydrophobic region: aa 95-132 truncated) failed to rescue [Prnp.sup.-/-] cells from apoptosis.

Caption: Fig. 5. Summarized schematic representation of phenotypes of [Prnp.sup.-/-] mice. Under the stressful conditions, [Prnp.sup.-/-] mice showed various phenotypes. Detail of these phenotypes is summarized in reference 119. In vivo studies have demonstrated that [Prnp.sup.-/-] mice are more prone to seizure, depression, and induction of epilepsy as well as experience extensive cerebral damage following ischemic challenge or viral infection. In experimental autoimmune encephalomyelitis, [Prnp.sup.-/-] mice reportedly have a more aggressive disease onset and clinical improvement during chronic phase than wild-type mice. [Prnp.sup.-/-] mice demonstrated significantly greater increase in blood glucose concentration after intraperitoneal injection of glucose than wild-type mice. In mice given oral dextran sulfate, Pr[P.sup.C] indicated a potential protective role against inflammatory bowel disease.

Caption: Fig. 6. Pr[P.sup.C]-based lipid raft signaling platform. The co-receptors LRP1 and mGluR5 clusters with Pr[P.sup.C] upon A[BETA]O binding leads to activation of Fyn kinase. Fyn kinase phosphorylates NMDAR and tau. Subsequently tau becomes pTyr18 (phosphor-Tyr-18). NMDAR becomes pTyr1482NR2 (phosphor-Tyr-1482 NMDAR). NMDAR and tau (pTyr18) then induce synaptic impairment and neurodegeneration.
Table 1. Conventional virus-induced slow infections of man

Virus                        Diseases

                                  RNA viruses


Poliovirus                 Chronic meningoencephalitis
                           in immunodeficient patient
Enterovirus                Virus-induced diabetes,
                           subacute encephalitis
Echovirus Chronic
                           meningoencephalitis in
                           immunodeficient patient


Measles                    Sub acute sclerosing


Rubella                      Progressive
                           congenital rubella


Tick-borne encephalitis    Epilepsia partialis
                           continua (Kozhevnikov's
                           epilepsy),RSSE, progressive
                           bulbar palsy in Russia
Hepatitis C                  Chronic hepatitis C
                           (non-A, non-B hepatitis)


Lentivirus HIV-1, HIV-2    HIV neuromyeloencephalopathy

Oncovirus HTLV-1, HTLV-2   Adult T-cell leukemia and mycosis
                           fungoides, HAM/TSP
                           neuromyeloencephalopathy, Jamaican
                           Neuropathy, tropical spastic paraparesis

Rabies lyssavirus          Rabies encephalitis

                                  DNA viruses


Adenovirus 7 and 32        Subacute encephalitis


JC virus                   Progressive multifocal
SV40                       Progressive multifocal


Hepatitis B                Homologous serum jaundice


Herpes simplex             Sub acute encephalitis
Cytomegalovirus            Cytomegalovirus brain
Epstein-Barr virus         Chronic infectious mononucleosis
Varicella-Zoster virus     Herpes zoster (sub acute encephalitis)
Herpes saimiri (B-virus)   Chronic herpes B encephalitis

Table 2. Transmissible spongiform encephalopathies with mutations
responsible for inherited genetic forms of the disease

In humans


Creutzfeldt-Jakob disease

  Iatrogenic (human growth hormone, pituitary gonadotropin, dura matter
    and corneal transplants, stereotactic electrodes)
  Familial (178asn, 200lys, 210ile, octapeptides inserts 2, 4-9)

Gerstmann-Straussler-Scheinker syndrome (GSS is only genetic,
  mutations are classified according to disease phenotype)

Fatal familial insomnia (178asn)

In Animals

Scrapie (sheep, goats, moufflon)
Transmissible mink encephalopathy
Chronic wasting disease (mule deer, elk)
Bovine spongiform encephalopathy
Exotic ungulate spongiform encephalopathy (nyala, gemsbok, Arabian
  oryx, greater kudu, eland)
Feline spongiform encephalopathy (cats, albino tiger, puma, cheetah)
COPYRIGHT 2017 The Japan Academy
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2017 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Onodera, Takashi
Publication:Japan Academy Proceedings Series B: Physical and Biological Sciences
Article Type:Report
Date:Apr 1, 2017
Previous Article:Proceedings at the 1105th general meeting.
Next Article:Purinergic signaling in microglia in the pathogenesis of neuropathic pain.

Terms of use | Privacy policy | Copyright © 2020 Farlex, Inc. | Feedback | For webmasters