A simple microfluidic platform to study age-dependent protein abundance and localization changes in Saccharomyces cerevisiae
Authors:Margarita Cabrera1,†, Daniele Novarina1, Irina L. Rempel1, Liesbeth M. Veenhoff1, and Michael Chang1
doi: 10.15698/mic2017.05.573
Volume 4, pp. 169 to 174, published 13/04/2017.
1 European Research Institute for the Biology of Ageing, University of Groningen, University Medical Center Groningen, 9713 AV Groningen, the Netherlands.
† Present address: Department of Experimental and Health Sciences, Pompeu Fabra University, 08003 Barcelona, Spain.
Keywords:
yeast, replicative ageing, microfluidics, CellASIC, protein abundance, protein localization.
Corresponding Author(s):
Conflict of interest statement:
The authors declare no conflict of interest.
Please cite this article as:
Margarita Cabrera, Daniele Novarina, Irina L. Rempel, Liesbeth M. Veenhoff, and Michael Chang (2017). A simple microfluidic platform to study age-dependent protein abundance and localization changes in Saccharomyces cerevisiae Microbial Cell 4(5): 169-174. doi: 10.15698/mic2017.05.573
© 2017 Cabrera et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
The budding yeast Saccharomyces cerevisiae divides asymmetrically, with a smaller daughter cell emerging from its larger mother cell. While the daughter lineage is immortal, mother cells age with each cell division and have a finite lifespan. The replicative ageing of the yeast mother cell has been used as a model to study the ageing of mitotically active human cells. Several microfluidic platforms, which use fluid flow to selectively remove daughter cells, have recently been developed that can monitor cell physiology as mother cells age. However, these platforms are not trivial to set up and users often require many hours of training. In this study, we have developed a simple system, which combines a commercially available microfluidic platform (the CellASIC ONIX Microfluidic Platform) and a genetic tool to prevent the proliferation of daughter cells (the Mother Enrichment Program), to monitor protein abundance and localization changes during approximately the first half of the yeast replicative lifespan. We validated our system by observing known age-dependent changes, such as decreased Sir2 abundance, and have identified a protein with a previously unknown age-dependent change in localization.