Abstract:
:Autophagy is a catabolic process that is crucial for cellular homeostasis and adaptive response to changing environments. Importantly, autophagy has been shown to be induced in many longevity-associated scenarios and to be required to maintain lifespan extension. Notably, autophagy is a highly conserved cellular process among eukaryotes, and the yeast Saccharomyces cerevisiae has become a universal model system for unraveling the molecular machinery underlying autophagic mechanisms. Here, we discuss different protocols to monitor survival and autophagy of yeast cells upon chronological aging. These include the use of propidium iodide to assess the loss of cell membrane integrity, as well as clonogenic assays to directly determine survival rates. Additionally, we describe methods to quantify autophagic flux, including the alkaline phosphatase activity or the GFP liberation assays, which measure the delivery of autophagosomal cargo to the vacuole. In sum, we have recapped established protocols used to evaluate a link between lifespan extension and autophagy in yeast.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Kainz K,Tadic J,Zimmermann A,Pendl T,Carmona-Gutierrez D,Ruckenstuhl C,Eisenberg T,Madeo Fdoi
10.1016/bs.mie.2016.09.086subject
Has Abstractpub_date
2017-01-01 00:00:00pages
367-394eissn
0076-6879issn
1557-7988pii
S0076-6879(16)30359-7journal_volume
588pub_type
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