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. 2009 Aug 12;29(32):10072-80.
doi: 10.1523/JNEUROSCI.2542-09.2009.

A New Transgenic Mouse Model of Gerstmann-Straussler-Scheinker Syndrome Caused by the A117V Mutation of PRNP

Wenbin Yang  1 , Julie Cook, Benjamin Rassbach, Azucena Lemus, Stephen J DeArmond, James A Mastrianni
Affiliations

A New Transgenic Mouse Model of Gerstmann-Straussler-Scheinker Syndrome Caused by the A117V Mutation of PRNP

Wenbin Yang et al. J Neurosci. .

Abstract

Gerstmann-Sträussler-Scheinker syndrome (GSS) is a genetic prion disease typified clinically by the development of progressive ataxia and dementia, and histopathologically by the presence of prion protein (PrP) amyloid plaques in the CNS, especially within the cerebellum. Several mutations of the PrP gene (PRNP) are associated with GSS, but only the P102L mutation has been convincingly modeled in transgenic (Tg) mice. To determine whether other mutations carry specific GSS phenotypic information, we constructed Tg mice that express PrP carrying the mouse homolog of the GSS-associated A117V mutation. Tg(A116V) mice express approximately six times the endogenous levels of PrP, develop progressive ataxia by approximately 140 d, and die by approximately 170 d. Compared with a mouse model of transmissible Creutzfeldt-Jakob disease (CJD), the ataxia of Tg(A116V) mice is more prominent, and the course of disease is more protracted, paralleling that observed in human disease. Neuropathology includes mild scattered vacuolation and prominent, mainly cerebellar localized, thioflavin S-positive PrP plaques comprised of full-length PrP(A116V). In some mice, more prominent vacuolation or a noncerebellar distribution of PrP plaques was evident, suggesting some variability in phenotype. The biophysical properties of PrP from Tg(A116V) mice and human GSS(A117V) revealed a similarly low fraction of insoluble PrP and a weakly protease-resistant approximately 13 kDa midspan PrP fragment, not observed in CJD. Overall, Tg(A116V) mice recapitulate many clinicopathologic features of GSS(A117V) that are distinct from CJD, supporting PrP(A116V) to carry specific phenotypic information. The occasional variation in histopathology they exhibit may shed light on a similar observation in human GSS(A117V).

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Figures

Figure 1.
Figure 1.
PrPA116V in Tg(A116V) mice. A, Western blot of brain homogenates from Tg(A116V) and WT mice displays a similar pattern and proportion of PrP glycoforms. B, Serially diluted whole-brain homogenates from 30-d-old asymptomatic and 150-d-old symptomatic Tg(A116V) mice, compared with 150 d-old WT mice. C, Plot of densitometric quantification of dot blot. At a young age, Tg(A116V) mice express approximately six times the normal level of PrPC. When symptomatic, the total level is approximately eight times that of WT mice, suggesting accumulation of PrP. The image was processed on a Bio-Rad XRS document imager, and Quantity One software (Bio-Rad) was applied to calculate the relative density against the dilution factor. Anti-mouse PrP D13 F(ab) antibody was used in each case.
Figure 2.
Figure 2.
Characteristics of spontaneous disease in Tg(A116V) mice. A, Kaplan–Meier survival curve of the rate of death plotted as a function of age in 15 Tg(A116V) mice. B, Mean age at symptom onset (in days) of 15 Tg(A116V) mice (onset, 148 ± 4 d) compared with an incubation period of 160 ± 5 d in 6 Tg(HuPrP)Prnpo/o mice that were intracerebrally inoculated with CJD(129MM1) brain homogenate. C, Mean age at death (in days) of Tg(A116V) mice was 176 ± 3 d, compared with the time to death from inoculation of 170 ± 4 d in CJD-inoculated Tg(HuPrP)Prnpo/o mice. D, Duration of disease in Tg(A116V) mice was 30 ± 3 d, compared with a very rapid 9 ± 1 d, in CJD-inoculated Tg(HuPrP)Prnpo/o mice (p < 0.001, Student's t test).
Figure 3.
Figure 3.
Neuropathological characteristics of Tg(A116V) mice. A, H&E stain shows sparse numbers of vacuoles in the hippocampus (CA1). B, PrP IHC with R2 antibody shows two PrP-positive amyloid plaques surrounded by vacuoles (florid plaques) in the hippocampus. C, GFAP IHC shows a moderate degree of reactive astrocytic gliosis in the hippocampus. D, PrP IHC (R2 antibody) shows PrP-positive amyloid plaques in the granule cell layer of the cerebellar cortex, as well as smaller punctate PrP-immunopositive deposits in both the molecular layer (M) and granular cell layer (G). E, Thioflavin S binds to the PrP plaques and displays fluorescence under UV light. F, GFAP IHC shows intense radial gliosis in the molecular layer and astrocytic gliosis in the granule cell layer. Scale bars: (in A) A, C, F, 50 μm; (in B) B, D, E, 50 μm.
Figure 4.
Figure 4.
Regional vacuolation of Tg(A116V) mice. A, Mean vacuolation scores from eight brain regions of seven symptomatic Tg(A116V) mice. NC, Neocortex; Hp, hippocampus; Th, thalamus; Hy, hypothalamus; Cd, caudate nucleus; pons; cbm, cerebellum molecular layer; cbg, cerebellum granular layer. The scoring system is described in Materials and Methods. The dashed line represents the limit at which vacuolation is reliably detected (score, 3). Error bars indicate SEM. B, The individual vacuolation scores of each of seven mice are plotted. The majority of scores were similarly low among the group, with the exception of mouse B, which displayed much higher levels in thalamus and caudate. Mouse F also had more prominent vacuolation in the thalamus, compared with the remainder of the group.
Figure 5.
Figure 5.
PrP amyloid plaques in Tg(A116V) mice are composed of full-length PrPA116V. IHC was performed using four PrP antibodies that recognize four distinct regions of mature PrP23-230, as indicated. All plaques are from the granule cell layer of the cerebellar cortex. “No Primary Ab” signifies all of the IHC steps were performed except incubation of the slide with a primary anti-PrP antibody.
Figure 6.
Figure 6.
PrPA116V insolubility compares with human GSS(A117V). A, Supernatant (S) and pellet (P) fractions prepared from brain homogenates of 150-d-old WT mice, asymptomatic 30-d-old and symptomatic 160-d-old Tg(A116V) mice, and a terminally ill scrapie-infected WT mouse (RML isolate). B, The solubility profiles of PrP from homogenates prepared from normal human brain (CTL), GSS(A117V), and sporadic CJD, are compared. Brain homogenates prepared as 10% w/v in lysis buffer and centrifuged at 100,000 × g at 4°C for 1 h to separate insoluble PrP, as described in Materials and Methods.
Figure 7.
Figure 7.
A ∼13 kDa rPrPSc fragment is detected in symptomatic Tg(A116V) mice and in GSS(A117V). PK titration of brain homogenates from a 60-d-old asymptomatic Tg(A116V) mouse (A), a 160-d-old symptomatic Tg(A116V) mouse (B), and human GSS(A117V) (C). Brain homogenates were digested with indicated concentrations of PK for 1 h at 37°C. Membranes were probed with D13 for MoPrP detection, and 3F4 to detect human PrP, as labeled. The open arrowheads indicate the ∼13 kDa fragments. A single fragment is detected in Tg(A116V) mice, whereas GSS(A117V) displayed a very faint ∼15 kDa fragment and a smaller doublet of ∼13 kDa.
Figure 8.
Figure 8.
Mapping endogenous cleavage products of Tg(A116V). Brain homogenates from an uninfected WT mouse (WT), a clinically sick WT mouse inoculated with mouse-adapted RML scrapie prions (RML), and a symptomatic (160-d-old) Tg(A116V) mouse (Tg), before (−) and after (+) deglycosylation with PNGase F, separated by SDS-PAGE on a 16% gel and probed separately with D13 or D18 antibodies. C1 and C2 are endogenous degradation products of PrPA116V that differ from typical scrapie.
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References

    1. Bessen RA, Marsh RF. Biochemical and physical properties of the prion protein from two strains of the transmissible mink encephalopathy agent. J Virol. 1992;66:2096–2101. - PMC - PubMed
    1. Brown P, Gibbs CJ, Jr, Rodgers-Johnson P, Asher DM, Sulima MP, Bacote A, Goldfarb LG, Gajdusek DC. Human spongiform encephalopathy: the National Institutes of Health series of 300 cases of experimentally transmitted disease. Ann Neurol. 1994;35:513–529. - PubMed
    1. Büeler H, Fischer M, Lang Y, Bluethmann H, Lipp HP, DeArmond SJ, Prusiner SB, Aguet M, Weissmann C. Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein. Nature. 1992;356:577–582. - PubMed
    1. Carlson GA, Ebeling C, Yang SL, Telling G, Torchia M, Groth D, Westaway D, DeArmond SJ, Prusiner SB. Prion isolate specified allotypic interactions between the cellular and scrapie prion proteins in congenic and transgenic mice. Proc Natl Acad Sci U S A. 1994;91:5690–5694. - PMC - PubMed
    1. Chen SG, Teplow DB, Parchi P, Teller JK, Gambetti P, Autilio-Gambetti L. Truncated forms of the human prion protein in normal brain and in prion diseases. J Biol Chem. 1995;270:19173–19180. - PubMed

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