HAL Id: hal-02741955
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A biochemical approach to evaluate the contribution to pathological prion protein formation from the 222K PrP
variant in scrapie positive goats
Maria Mazza, Chiara Guglielmetti, Francesco Ingravalle, Jan P. M. Langeveld, Loukia V. Ekateriniadou, Olivier Andréoletti, Pier Luigi Acutis
To cite this version:
Maria Mazza, Chiara Guglielmetti, Francesco Ingravalle, Jan P. M. Langeveld, Loukia V. Ekaterini- adou, et al.. A biochemical approach to evaluate the contribution to pathological prion protein for- mation from the 222K PrP variant in scrapie positive goats. Prion 2015, May 2015, Fort Collins, Colorado, United States. Taylor & Francis, Prion, 9 (suppl. 1), 99 p., 2015, Prion 2015 Poster Abstracts. �hal-02741955�
oligomeric A-Beta peptide has a major binding site between the alpha- and beta-sites of PrPC.
b-cleavage of PrPChas previously been sug- gested to occur through reactive oxygen spe- cies generated by the binding of copper to the PrP OR. We have recently reported gen- eration of a novel Prnp allele, “S3” which has PrPC OR conformationally-locked in a compact arrangement that favors binding of one copper ion per OR. Interestingly, this allele resulted in overproduction of C2 PrP in Tg.S3 mice and transfected RK13 cells, but not N2a and SMB cells. These observa- tions, in conjunction with chelator studies, imply that b-processing of PrPC may occur by a mechanism distinct from metal cata- lyzed hydrolysis reported in vitro. In this current study, using mass spectroscopy anal- ysis of mouse brain S3 and Wt PrPC, we have identified the “P1” amino acid residue for b-cleavage in vivo. This information, as well as studies with protease inhibitors, has narrowed the possible identities for a b-site protease. As alpha- versus beta-processing of PrPC can lead to benign outcomes in the presence of PrPSc or oligomeric A-Beta, our findings may provide useful insights into the pathogenesis of both prion and Alzheimer’s disease.
P.32: A biochemical approach to evaluate the contribution to pathological
prion protein formation from the 222K PrP variant in scrapie positive goats
Maria Mazza
1, Chiara Guglielmetti
1, Francesco Ingravalle
2, Jan PM Langeveld
3,
Loukia V Ekateriniadou
4,
Olivier Andr eoletti
5, and Pier Luigi Acutis
11Genetics and Immunobiochemistry Laboratory;
Department of Neurosciences; Istituto Zooprofilattico Sperimentale del Piemonte, Liguria
e Valle d’Aosta, Turin, Italy;2Biostatistic;
Epidemiology and Risk Analysis Unit; Istituto Zooprofilattico Sperimentale del Piemonte, Liguria
e Valle d’Aosta, Turin, Italy;3Central Institute for Animal Disease Control (CIDC-Lelystad), Lelystad,
The Netherlands;4National Agricultural Research Foundation; Veterinary Research Institute, Thessaloniki, Greece;5UMR INRA ENVT 1225;
Interactions HOtes Agents Pathog^ enes;
Ecole Nationale Veterinaire de Toulouse, Toulouse, France
Lysine (K) at position 222 of goat PrP is asso- ciated with resistance to classical scrapie, yet few natural cases of scrapie have been found in Q/K goats and intracerebral experimental trans- mission was successful in Q/K and K/K goats after long incubation periods. Previous Western Blot (WB) studies performed by different mono- clonal antibodies (mAbs) revealed an inhibition by K222 on the binding of mAb F99/97.6.1 to goat PrP. Thus a WB method, based on the ratio between the signal intensity given by mAbs F99/97.6.1 and SAF84, was developed to distin- guish goats exhibiting PrP encoded by the geno- types 222Q/Q, 222Q/K and 222K/K. Here we applied this approach to investigate the contribu- tion to PrPScformation given by the 222K vari- ant in scrapie positive goats. WB analyses were performed on the PrPCor PrPSCextracted from the brains of negative and scrapie positive goat samples (natural or experimental), bearing dif- ferent genotypes at position 222. Samples were analyzed in triplicate and signals revealed by F99/97.6.1 and SAF84 were quantified. A descriptive statistical analysis was performed.
S28 Prion 2015 Poster Abstracts
No signals were detected by F99/97.6.1 in any of the 222K/K goat samples. The ratio of the optical densities revealed that the positive 222Q/K goats had a similar reactivity to 222Q/
Q samples. Halved values of the ratio were present in negative goats with 222Q/K. The sta- tistical analysis confirmed these differences.
The obtained results showed that in heterozy- gous animals PrPSc seems to be formed nearly totally by the Q222 variant thus confirming the higher resistance to convertibility of K222 PrP variant.
P.33: Genetic modulation of atypical scrapie transmissions in transgenic
mouse models
Hae-Eun Kang
1, Sehun Kim
1, April Zander
1, Thierry Baron
2, and
Glenn Telling
11Prion Research Center (PRC) and the Department of Microbiology; Immunology and Pathology;
Colorado State University; Fort Collins, CO USA;
2Agence Fran¸caise de Securite Sanitaire des Aliments; Lyon, France
Atypical scrapie, a recently recognized and surprisingly prevalent prion disease of sheep, is an example of an emerging prion disease of uncertain origin and species tropism. Since its discovery in 1998, precautionary testing have revealed surprising numbers of atypical scrapie cases in Europe. In 2007, a sheep from a flock in Wyoming tested positive for atypical scrapie.
Atypical and classical scrapie differ with bio- chemical features of the protease-resistant form of the prion protein (PrPSc), and neuropathol- ogy. Also, atypical scrapie occurs in sheep with PRNPgenotypes usually associated with resis- tance to classical scrapie, and sheep with atypi- cal scrapie are more likely to express phenylalanine (F) than leucine (L) at codon 141. The origin and host range of atypical scra- pie are unclear. To address these issues, we generated transgenic mice expressing Ovine PrP with phenylalanine at the residue 141, Tg (OvPrP-F141). We produced 2 sublines: the brains of the Tg(OvPrP-F141)H express PrPC
»5-fold higher than wild type mice, while Tg (OvPrP-F141)L mice express at »2-fold. Tg (OvPrP-F141)H spontaneously developed trun- cal ataxia and limb paresis at a mean age of 70 d. Remarkably, PrPSc accumulated in brains and muscles of sick mice. Furthermore, we inoculated the Tg(OvPrP-F141)L, Tg(OvPrP- A136), and Tg(OvPrP-V136) mice with 6 atyp- ical scrapie isolates to determine the effects of the F141 genotype on atypical scrapie propaga- tion. Only Tg(OvPrP-F141)L developed clini- cal signs and the brain samples showed typical biochemical signature of atypical scrapie. Our findings suggest that atypical scrapie may have spontaneously originated and subsequently propagated and evolved by transmission in sheep with susceptible genotypes.
P.34: Preliminary study of Alzheimer’s disease transmission to bank vole
Guido Di Donato
1, Geraldina Riccardi
1, Claudia D’Agostino
1, Flavio Torriani
1,
Maurizio Pocchiari
2, Romolo Nonno
1, Umberto Agrimi
1, and
Michele Angelo Di Bari
11Department of Food Safety and Veterinary Public Health Istituto Superiore di Sanita, Rome, Italy;
2Department of Cellular Biology and Neuroscience;
Istituto Superiore di Sanita, Rome, Italy
Extensive experimental findings indicate that prion-like mechanisms underly the patho- genesis of Alzheimer disease (AD). Transgenic mice have been pivotal for investigating prion- like mechanisms in AD, still these models have not been able so far to recapitulate the complex clinical-pathological features of AD. Here we aimed at investigating the potential of bank vole, a wild-type rodent highly susceptible to prions, in reproducing AD pathology upon experimental inoculation.
Voles were intracerebrally inoculated with brain homogenate from a familial AD patient.
Animals were examined for the appearance of neurological signs until the end of experiment (800 d post-inoculation, d.p.i.). Brains were studied by immunohistochemistry for pTau
Prion 2015 Poster Abstracts S29
