title: Thioflavin-S staining coupled to flow cytometry. A screening tool to detect in vivo protein aggregation.
abstract: Amyloid deposits are associated with an increasing number of human disorders, including Alzheimer's and Parkinson's diseases. Recent studies provide compelling evidence for the existence of amyloid-like conformations in the insoluble bacterial inclusion bodies (IBs) produced during the recombinant expression of amyloidogenic proteins. This makes prokaryotic cells a physiologically relevant system to study the mechanisms of in vivo amyloid deposition. We show here that the application of flow cytometry to detect Thioflavin-S (Th-S) fluorescence provides a fast, robust, quantitative, non-invasive method to screen for the presence of in vivo intracellular amyloid-like aggregates in bacteria, with potential application in the analysis of the impact of genetic mutations or chemical compounds on the aggregation of disease-associated polypeptides.
authors.affiliation: Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, 08193-Bellaterra, Barcelona, Spain.
authors.forename: Alba
authors.initials: A
authors.lastname: Espargaró
authors.forename: Raimon
authors.initials: R
authors.lastname: Sabate
authors.forename: Salvador
authors.initials: S
authors.lastname: Ventura
mesh_terms: D000682:Amyloid / Q000378:metabolism; D052160:Benzothiazoles; D005434:Flow Cytometry / Q000379:methods*; D002479:Inclusion Bodies / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry*
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D011506:Proteins; D013844:Thiazoles; C009462:thioflavin T