Neurogenerative Diseases - 19

such as seeding assays (Thioflavin T assay detailed
prior), sedimentation assays to show aggregated
material in denaturing and non-denaturing gel
electrophoresis, transmission and atomic force EM
techniques, circular dichroism spectroscopy and
Fourier-transform infrared spectroscopy, there has
been no work identifying the degree of solvation
that occurs during the sonication process.
When fibrils are made, they are essentially 100%
desolvated, meaning they are a suspension rather
than a solution. This means that the fibrils can be
spun off easily using centrifugation, and as they are
insoluble, they are able to clump together depending
Figure 3. Thioflavin T emission curves show increased
fluorescence (correlated to alpha-synuclein protein aggregation)
over time when 10 µM of Type 1 alpha-synuclein PFFs (SPR-322)
are combined with 100 µM of alpha-synuclein monomer (SPR321), as compared to Type 1 alpha-synuclein PFFs (SPR-322) alone
and alpha-synuclein monomer (SPR-321) alone.

Figure 2. Alpha-synuclein and tau PFFs induce endogenous
phosphorylation and aggregation of their respective proteins
in vitro and in vivo. (A) ICC analysis of rat primary neurons treated
with Type 1 human alpha-synuclein PFFs (SPR-322) shows Lewy
body pathology 15 days after treatment. Blue: Hoechst. Green:
pSer129. (B) ICC analysis of ATTO633-labelled Type 1 human
alpha-synuclein PFFs (SPR-322) added to mouse primary cells
shows seeding and transport within the soma and neurites within
24 hours. Image courtesy of Cellectricon. (C) IHC analysis of P301L
mouse brain injected with K18 tau P301L PFFs (SPR-330) shows
AT8-positive tau inclusions in the hippocampus 9 weeks postinjection. Image courtesy of ReMYND N.V. (D) IHC analysis of rat
brain injected with Type 1 mouse alpha-synuclein PFFs (SPR-324)
shows Lewy body pathology 30 days post-injection.

Figure 4. TEM of various human alpha-synuclein PFF and oligomer
preparations. (A) Type 1 PFFs (SPR-322). (B) Type 2 PFFs (SPR-317).
(C) Type 3 PFFs (SPR-448). (D) Dopamine-stabilized oligomers
(SPR-466). For more information on fibril types see stressmarq.
com/SNCA.

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Neurogenerative Diseases

Table of Contents for the Digital Edition of Neurogenerative Diseases
