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Open AccessArticle

The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach

1
Department of Physiological Sciences, Stellenbosch University, Stellenbosch 7602, South Africa
2
Department of Biochemistry, Stellenbosch University, Stellenbosch 7602, South Africa
3
Department of Physics, Stellenbosch University, Stellenbosch 7602, South Africa
*
Author to whom correspondence should be addressed.
Cells 2018, 7(8), 94; https://doi.org/10.3390/cells7080094
Received: 3 July 2018 / Revised: 31 July 2018 / Accepted: 31 July 2018 / Published: 3 August 2018
(This article belongs to the Special Issue Autophagy in Age-Related Human Diseases)
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. View Full-Text
Keywords: autophagic flux; vesicle dynamics; autophagosome; autolysosome; lysosome; pool size; velocity; displacement; micro-pattern autophagic flux; vesicle dynamics; autophagosome; autolysosome; lysosome; pool size; velocity; displacement; micro-pattern
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Du Toit, A.; De Wet, S.; Hofmeyr, J.-H.S.; Müller-Nedebock, K.K.; Loos, B. The Precision Control of Autophagic Flux and Vesicle Dynamics—A Micropattern Approach. Cells 2018, 7, 94.

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