eBook: Advancing Neuroscience Research - 16

Sonication of Pre-formed Fibrils
for Neurodegenerative Disease
Research: What, Why, How and
Best Practices
Sonication is an important step for preparing alpha-synuclein PFFs
for use in vitro and in vivo.
Alzheimer's and Parkinson's are the two most
common neurodegenerative diseases. They are
pathologically characterized by the misfolding and
aggregation of various proteins including alpha-synuclein,
tau, and amyloid-beta. These diseases are
incurable and becoming increasingly prevalent as
populations age.
Most basic disease research requires the use of
models or systems that generate the disease state
or a valid proximity of it. This is true for Alzheimer's
and Parkinson's diseases as well. Transgenic animals
can be used but will typically have long growth or
maturity times before they can be used-measured
in many months or more. Their early survival
post-birth is also not a given. One solution to this
problem has been the generation of recombinant
fibrillar protein constructs that, once injected into
a host, mimic what happens in these disease states,
generating a similar pathology to what is associated
with those diseases. Furthermore, when these
constructs are used, the pathology can be fully generated
within 30 days (in some cases), providing a
clear time advantage over, for instance, transgenic
animals that overexpress certain proteins and may
take 6-18 months to mature.
Neuroscientists have achieved remarkable progress
in studying neurodegenerative diseases. Although
challenges and barriers remain, scientists
are now able to generate disease models in a short
period of time by using these recombinant fibrillar
protein constructs, also called pre-formed fibrils,
or PFFs for short, which act as " seeds " to induce
protein aggregation. This " seeding " activity of PFFs
means that they can recruit soluble endogenous
proteins to form aggregates and ultimately induce
neurodegenerative pathology.
Pre-formed fibrils can be used in cell culture or
animal brains, where they seed and dramatically
accelerate the aggregation of monomers. This can
be seen in vitro using thioflavin T as an indicator,
where an increase in fluorescence shows the development
of beta sheets-an integral element of the
aggregated forms. Adding PFFs to primary neurons
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