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Tytuł pozycji:

Comparison of size distribution and (Pro249-Ser258) epitope exposure in in vitro and in vivo derived Tau fibrils

Tytuł :
Comparison of size distribution and (Pro249-Ser258) epitope exposure in in vitro and in vivo derived Tau fibrils
Autorzy :
André Marreiro
Kristof Van Kolen
Cristiano Sousa
Liesbet Temmerman
Bruno Vasconcelos
Rosa Crespo-Rodriguez
Jan R. T. van Weering
Debby Van Dam
Peter P. De Deyn
Adrian Apetri
Liliane Schoofs
Marc H. Mercken
Pokaż więcej
Temat :
Alzheimer’s disease
Tau
Tau aggregation
Aggregation
Seeding
Cytology
QH573-671
Źródło :
BMC Molecular and Cell Biology, Vol 21, Iss 1, Pp 1-14 (2020)
Wydawca :
BMC, 2020.
Rok publikacji :
2020
Kolekcja :
LCC:Cytology
Typ dokumentu :
article
Opis pliku :
electronic resource
Język :
English
ISSN :
2661-8850
Relacje :
http://link.springer.com/article/10.1186/s12860-020-00320-y; https://doaj.org/toc/2661-8850
DOI :
10.1186/s12860-020-00320-y
Dostęp URL :
https://doaj.org/article/b4b1144feeae41369a9bef997ed0bec5
Numer akcesji :
edsdoj.b4b1144feeae41369a9bef997ed0bec5
Czasopismo naukowe
Abstract Background Although several studies demonstrate prion-like properties of Tau fibrils, the effect of size in the seeding capacity of these aggregates is not fully understood. The aim of this study is to characterize Tau seeds by their size and seeding capacity. Methods Tau aggregates were isolated from postmortem AD brain tissue and separated from low molecular weight species by sucrose gradient ultracentrifugation. Biochemical characterization of the different fractions was done by non-reducing Western blotting and aggregate-specific immuno-assays using in house developed anti-Tau monoclonal antibodies, including PT76 which binds to an epitope close to the microtubule-binding domain and, hence, also to K18. Seeding efficiency was then assessed in HEK293 cells expressing K18 FRET sensors. Results We observed that upon sonication of Tau aggregates different size-distributed tau aggregates are obtained. In biochemical assays, these forms show higher signals than the non-sonicated material in some aggregation-specific Tau assays. This could be explained by an increased epitope exposure of the smaller aggregates created by the sonication. By analyzing human brain derived and recombinant (K18) Tau aggregates in a cellular FRET assay, it was observed that, in the absence of transfection reagent, sonicated aggregates showed higher aggregation induction. Preparations also showed altered profiles on native PAGE upon sonication and we could further separate different aggregate species based on their molecular weight via sucrose gradients. Conclusions This study further elucidates the molecular properties regarding relative aggregate size and seeding efficiency of sonicated vs. non-sonicated high molecular weight Tau species. This information will provide a better knowledge on how sonication, a commonly used technique in the field of study of Tau aggregation, impacts the aggregates. In addition, the description of PT76-based aggregation specific assay is a valuable tool to quantify K18 and human AD Tau fibrils.
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