The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach
Autophagy failure is implicated in age-related human disease. A decrease in the rate of protein degradation through the entire autophagy pathway, i.e., autophagic flux, has been associated with the onset of cellular proteotoxity and cell death. Although the precision control of autophagy as a pharma...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2018-08-01
|
Series: | Cells |
Subjects: | |
Online Access: | http://www.mdpi.com/2073-4409/7/8/94 |
id |
doaj-3bb8e627e0d74c63a1acbab790afa276 |
---|---|
record_format |
Article |
spelling |
doaj-3bb8e627e0d74c63a1acbab790afa2762020-11-25T01:01:53ZengMDPI AGCells2073-44092018-08-01789410.3390/cells7080094cells7080094The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern ApproachAndré du Toit0Sholto De Wet1Jan-Hendrik S. Hofmeyr2Kristian K. Müller-Nedebock3Ben Loos4Department of Physiological Sciences, Stellenbosch University, Stellenbosch 7602, South AfricaDepartment of Physiological Sciences, Stellenbosch University, Stellenbosch 7602, South AfricaDepartment of Biochemistry, Stellenbosch University, Stellenbosch 7602, South AfricaDepartment of Physics, Stellenbosch University, Stellenbosch 7602, South AfricaDepartment of Physiological Sciences, Stellenbosch University, Stellenbosch 7602, South AfricaAutophagy failure is implicated in age-related human disease. A decrease in the rate of protein degradation through the entire autophagy pathway, i.e., autophagic flux, has been associated with the onset of cellular proteotoxity and cell death. Although the precision control of autophagy as a pharmacological intervention has received major attention, mammalian model systems that enable a dissection of the relationship between autophagic flux and pathway intermediate pool sizes remain largely underexplored. Here, we make use of a micropattern-based fluorescence life cell imaging approach, allowing a high degree of experimental control and cellular geometry constraints. By assessing two autophagy modulators in a system that achieves a similarly raised autophagic flux, we measure their impact on the pathway intermediate pool size, autophagosome velocity, and motion. Our results reveal a differential effect of autophagic flux enhancement on pathway intermediate pool sizes, velocities, and directionality of autophagosome motion, suggesting distinct control over autophagy function. These findings may be of importance for better understanding the fine-tuning autophagic activity and protein degradation proficiency in different cell and tissue types of age-associated pathologies.http://www.mdpi.com/2073-4409/7/8/94autophagic fluxvesicle dynamicsautophagosomeautolysosomelysosomepool sizevelocitydisplacementmicro-pattern |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
André du Toit Sholto De Wet Jan-Hendrik S. Hofmeyr Kristian K. Müller-Nedebock Ben Loos |
spellingShingle |
André du Toit Sholto De Wet Jan-Hendrik S. Hofmeyr Kristian K. Müller-Nedebock Ben Loos The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach Cells autophagic flux vesicle dynamics autophagosome autolysosome lysosome pool size velocity displacement micro-pattern |
author_facet |
André du Toit Sholto De Wet Jan-Hendrik S. Hofmeyr Kristian K. Müller-Nedebock Ben Loos |
author_sort |
André du Toit |
title |
The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach |
title_short |
The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach |
title_full |
The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach |
title_fullStr |
The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach |
title_full_unstemmed |
The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach |
title_sort |
precision control of autophagic flux and vesicle dynamics—a micropattern approach |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2018-08-01 |
description |
Autophagy failure is implicated in age-related human disease. A decrease in the rate of protein degradation through the entire autophagy pathway, i.e., autophagic flux, has been associated with the onset of cellular proteotoxity and cell death. Although the precision control of autophagy as a pharmacological intervention has received major attention, mammalian model systems that enable a dissection of the relationship between autophagic flux and pathway intermediate pool sizes remain largely underexplored. Here, we make use of a micropattern-based fluorescence life cell imaging approach, allowing a high degree of experimental control and cellular geometry constraints. By assessing two autophagy modulators in a system that achieves a similarly raised autophagic flux, we measure their impact on the pathway intermediate pool size, autophagosome velocity, and motion. Our results reveal a differential effect of autophagic flux enhancement on pathway intermediate pool sizes, velocities, and directionality of autophagosome motion, suggesting distinct control over autophagy function. These findings may be of importance for better understanding the fine-tuning autophagic activity and protein degradation proficiency in different cell and tissue types of age-associated pathologies. |
topic |
autophagic flux vesicle dynamics autophagosome autolysosome lysosome pool size velocity displacement micro-pattern |
url |
http://www.mdpi.com/2073-4409/7/8/94 |
work_keys_str_mv |
AT andredutoit theprecisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT sholtodewet theprecisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT janhendrikshofmeyr theprecisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT kristiankmullernedebock theprecisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT benloos theprecisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT andredutoit precisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT sholtodewet precisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT janhendrikshofmeyr precisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT kristiankmullernedebock precisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach AT benloos precisioncontrolofautophagicfluxandvesicledynamicsamicropatternapproach |
_version_ |
1725206961792745472 |