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In vivo amyloid aggregation kinetics tracked by time-lapse confocal microscopy in real-time.
- Source :
-
Biotechnology journal [Biotechnol J] 2016 Jan; Vol. 11 (1), pp. 172-7. Date of Electronic Publication: 2016 Jan 05. - Publication Year :
- 2016
-
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.<br /> (Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Subjects :
- Amyloid beta-Peptides biosynthesis
Amyloid beta-Peptides genetics
Escherichia coli metabolism
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Humans
Kinetics
Microscopy, Confocal methods
Protein Aggregates
Time-Lapse Imaging methods
Amyloid beta-Peptides chemistry
Escherichia coli genetics
Inclusion Bodies chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1860-7314
- Volume :
- 11
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Biotechnology journal
- Publication Type :
- Academic Journal
- Accession number :
- 26580000
- Full Text :
- https://doi.org/10.1002/biot.201500252