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Because parameters other than prrmary structure above may modulate spe-
cies barrier effects (see earher; and ref 31), recommendattons listed here
enhance, but do not guarantee, success m constructing mice sensitive to heter-
ologous prtons. Also tt 1s worth notmg that TgPrP mice may have an mtrmstc
limit m then susceptrbtlity to prton disease, corresponding to an mcubatton
time of ca. 50 d (21,40,58): This behavior 1s exemplified by moculatron of
Sc237 prtons mto Tg(SHaPrP)7+˜+ mice wtthm the PrnpO™O background, and by
moculatton of mouse-adapted RML prrons mto Tg(Mo-PrP-A)B4053 mice
(18). Although these transgemc mice represent a considerable advance over
transmtsslons into nontransgenic hosts, tt should be borne m mmd that animal
btoassays ulttmately ˜111 be supplanted by faster (and most probably cheaper)
in vitro systems for the converston of PrPC to PrPSC (59,60)

Note Added in Proof
Tellmg et al. (Cell 83,79-90,1995) have documented an mhlbltory effect of
mouse PrPC on transmtsston of human prlons mto Tg(Hu) PrP mice The
259
Pnon “Species-Barrier” Effects

authors hypothesize that mteractlons between PrPC and a macromolecule
designated “protein X” provide an additional level of specificity m specles-
barrier effects.

Acknowledgments
I am indebted to George Carlson, Richard Rubenstem, and John Collmge
for their comments, and the Alzhelmer™s Assoclatlon of Ontario for support.
References
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Methods for Studying Prion Protein Amyloid
Fabrizio Tagliavini, Frances Prelli, Giorgio Giaccone,
Gianluigi Forloni, Mario Salmona, Pedro Piccardo,
Bernardino Ghetti, Blas Frangione, and Orso Bugiani


1. Introduction
Prion encephalopathies are neurodegenerative diseasescharacterized by the
accumulation of abnormal isoforms of the priori protein (PrP) and the deposi-
tion of PrP amyloid m the central nervous system (CNS) (1,2) The disease-
specific PrP molecules are distinguishable from their normal homologs by their
relative resistance to proteinase K digestion (1,2), they are thought to be
derived from protease-sensitive precursors by a posttranslational process that
mvolves a conformational change with a shift from a-helix to P-sheet structure
(3--5)
Amyloid formation occurs to the highest degree m Gerstmann-straussler-
Schemker (GSS) disease (Fig. 1), an autosomal dominant disorder that exhib-
its a wide spectrum of clmical presentations, mcludmg ataxia, spastic
paraparesis, parkmsomsm, and dementia, and is associated with variant PRNP
genotypes resultmg from the combmation of a pathogenic mutation with a com-
mon polymorphism (ATG + GTG, M/V) at codon 129 (Fig. 2) (6). To date,
point mutations at codons 102 (CCG + CTG, P + L) and 145 (TAT + TAG,
Y + stop) have been found on a M 129 allele, whereas mutations at codons 105
(CCA + CTA, P -+ L), 117 (GCG -+ GTG, A + V), 198 (TTC + TCC, F -+ S),
and 217 (CGG + CAG, Q -+ R) have been detected on a V129 allele (6).
The climcal variability of GSS diseaseis related to the distribution and extent
of amyloid deposition as well as the occurrence of associated lesions (6).
Umcentric and multicentric amyloid deposits may be found throughout the
CNS. In most cases they are particularly abundant m the cerebellar cortex,

From Methods in Molecular Medmne Prlon Diseases
EdIted by H Baker and R M Rldley Humana Press Inc , Totowa, NJ

265
Tagliavini et al.
266




Fig. 1. Amyloid deposits in the cerebellar cortex of a patient with GSS disease. (A)
Amyloid cores are labeled by an antiserum to a synthetic peptide homologous to resi-
dues 9&l 02 of PrP. (B) At the electron microscopic level, amyloid deposits are com-
posed of radially oriented, S-IO-nm diameter fibrils. Bars: (A), 50 urn; (B), 2 urn.


102105117 145 198 217




stop s R
L L v

Fig. 2. Schematic representation of human PrP showing the mutations associated
with GSS disease and PrP-CAA. The protein consists of 253 residues. A signal pep-
tide of 22 amino acids is cleaved at the N terminus during biosynthesis and a hydro-
phobic signal sequence of 23 amino acids is removed from the C terminus on addi-

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