Protease-Resistant Prions Selectively Decrease Shadoo Protein

The central event in prion diseases is the conformational conversion of the cellular prion protein (PrPC) into PrPSc, a partially protease-resistant and infectious conformer. However, the mechanism by which PrPSc causes neuronal dysfunction remains poorly understood. Levels of Shadoo (Sho), a protein that resembles the flexibly disordered N-terminal domain of PrPC, were found to be reduced in the brains of mice infected with the RML strain of prions [1], implying that Sho levels may reflect the presence of PrPSc in the brain. To test this hypothesis, we examined levels of Sho during prion infection using a variety of experimental systems. Sho protein levels were decreased in the brains of mice, hamsters, voles, and sheep infected with different natural and experimental prion strains. Furthermore, Sho levels were decreased in the brains of prion-infected, transgenic mice overexpressing Sho and in infected neuroblastoma cells. Time-course experiments revealed that Sho levels were inversely proportional to levels of protease-resistant PrPSc. Membrane anchoring and the N-terminal domain of PrP both influenced the inverse relationship between Sho and PrPSc. Although increased Sho levels had no discernible effect on prion replication in mice, we conclude that Sho is the first non-PrP marker specific for prion disease. Additional studies using this paradigm may provide insight into the cellular pathways and systems subverted by PrPSc during prion disease

Accuracy of customized abutment data superimposition according to the extent of scanning area

To evaluate the accuracy of superimposition of customized abutment library data onto scanned abutment data according to the extent of the scanning area. A patient model was fabricated by a 3D printer (Probo, DIO Implant), and a customized abutment was fabricated using a four-axis milling machine (ARUM 4X-100, Doowon). The customized abutment library data were generated using a laboratory scanner (E3, 3Shape) for superimposition after intraoral scanning. A cone-shaped structure was embedded into the library data at the center of the connection part. The customized abutment was placed on the model, and the model was scanned using a laboratory scanner to produce reference data. Three different test group datasets were generated using intraoral scanner and computer-aided design software: (1) fully scanned customized abutment; (2) insufficiently scanned proximal surface; and (3) insufficiently scanned margin, assuming challenging intraoral conditions. The library data were superimposed onto each test group; thereafter, the distance and angle between the reference and test group data were analyzed by using the embedded cone. Statistical analysis was performed using one-way analysis of variance followed by post hoc Tukey test for multiple comparisons. There were no statistically significant differences between the mean distance and angle of the test group data (with three different scanning areas) and the reference data. The superimposition technique can be used clinically, not only when the scan is complete, but also when the proximal surface and margin of the customized abutment have been scanned incompletely.</p

Disruption of Doppel prevents neurodegeneration in mice with extensive Prnp deletions

The Prnp gene encodes the cellular prion protein PrP(C). Removal of its ORF does not result in pathological phenotypes, but deletions extending into the upstream intron result in cerebellar degeneration, possibly because of ectopic cis-activation of the Prnd locus that encodes the PrP(C) homologue Doppel (Dpl). To test this hypothesis, we removed Prnd from Prnp(o/o) mice by transallelic meiotic recombination. Balanced loxP-mediated ablation yielded mice lacking both PrP(C) and Dpl (Prn(o/o)), which developed normally and showed unimpaired immune functions but suffered from male infertility. However, removal of the Prnd locus abolished cerebellar degeneration, proving that this phenotype is caused by Dpl upregulation. The absence of compound pathological phenotypes in Prn(o/o) mice suggests the existence of alternative compensatory mechanisms. Alternatively, Dpl and PrP(C) may exert distinct functions despite having partly overlapping expression profiles

Transgene-driven expression of the Doppel protein in Purkinje cells causes Purkinje cell degeneration and motor impairment

The Doppel (Dpl) and Prion (PrP) proteins show 25% sequence identity and share several structural features with only minor differences. Dpl shows a PrP-like fold of its C-terminal globular domain and lacks the flexible N-terminal tail. The physiological functions of both proteins are unknown. However, ubiquitous Dpl overexpression in the brain of PrP knockout mice correlated with ataxia and Purkinje cell degeneration in the cerebellum. Interestingly, a similar phenotype was reported in transgenic mice expressing an N-terminally truncated PrP (ΔPrP) in Purkinje cells by the L7 promoter (TgL7-ΔPrP). Coexpression of full-length PrP rescued both the neurological syndromes caused by either Dpl or ΔPrP. To evaluate whether the two proteins caused cerebellar neurodegeneration by the same mechanism, we generated transgenic mice selectively expressing Dpl in Purkinje cells by the same L7 promoter. Such mice showed ataxia and Purkinje cell loss that depended on the level of Dpl expression. Interestingly, the effects of high levels of Dpl were not counterbalanced by the presence of two Prnp alleles. By contrast, PrP coexpression was sufficient to abrogate motor impairment and to delay the neurodegenerative process caused by moderate level of Dpl. A similar situation was reported for the corresponding TgL7-ΔPrP mice supporting the concept that Dpl and ΔPrP cause cell death, possibly by interfering with a common signaling cascade essential for cell survival