title: In vivo amyloid aggregation kinetics tracked by time-lapse confocal microscopy in real-time.
abstract: Amyloid polymerization underlies an increasing number of human diseases. Despite this process having been studied extensively in vitro, aggregation is a difficult process to track in vivo due to methodological limitations and the slow kinetics of aggregation reactions in cells and tissues. Herein we exploit the amyloid properties of the inclusions bodies (IBs) formed by amyloidogenic proteins in bacteria to address the kinetics of in vivo amyloid aggregation. To this aim we used time-lapse confocal microscopy and a fusion of the amyloid-beta peptide (A β42) with a fluorescent reporter. This strategy allowed us to follow the intracellular kinetics of amyloid-like aggregation in real-time and to discriminate between variants exhibiting different in vivo aggregation propensity. Overall, the approach opens the possibility to assess the impact of point mutations as well as potential anti-aggregation drugs in the process of amyloid formation in living cells.
authors.affiliation: Departament de Bioquimica i Biologia Molecular, Universitat Autònoma de Barcelona, Barcelona, Spain.|Institut de Biotecnologia i Biomedicina (IBB), Universitat Autònoma de Barcelona, Bellatera, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Villar-Piqué
authors.affiliation: Departament de Fisicoquímica, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain.|Institut de Nanociència i Nanotecnologia (IN²UB), Spain.
authors.forename: Alba
authors.initials: A
authors.lastname: Espargaró
authors.affiliation: Departament de Bioquimica i Biologia Molecular, Universitat Autònoma de Barcelona, Barcelona, Spain. salvador.ventura@uab.es.|Institut de Biotecnologia i Biomedicina (IBB), Universitat Autònoma de Barcelona, Bellatera, Spain. salvador.ventura@uab.es.
authors.forename: Salvador
authors.initials: S
authors.lastname: Ventura
authors.affiliation: Departament de Fisicoquímica, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain. rsabate@ub.edu.|Institut de Nanociència i Nanotecnologia (IN²UB), Spain. rsabate@ub.edu.
authors.forename: Raimon
authors.initials: R
authors.lastname: Sabate
mesh_terms: D016229:Amyloid beta-Peptides / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D007700:Kinetics; D018613:Microscopy, Confocal / Q000379:methods; D066329:Protein Aggregates; D059008:Time-Lapse Imaging / Q000379:methods
chemical_list: D016229:Amyloid beta-Peptides; D066329:Protein Aggregates; D049452:Green Fluorescent Proteins
keywords: Aβ peptide; aggregation kinetics; amyloid aggregation; fibril formation; inclusion bodies