Guidelines for the use and interpretation of assays for monitoring autophagy

dc.contributor.author Klionsky, Daniel
dc.contributor.author Bassham, Diane
dc.contributor.author Amarnath, Shoba
dc.contributor.author Anantharam, Vellareddy
dc.contributor.author Bassham, Diane
dc.contributor.author Bellaire, Bryan
dc.contributor.department Biomedical Sciences
dc.contributor.department Veterinary Microbiology and Preventive Medicine
dc.contributor.department Genetics, Development and Cell Biology
dc.date 2018-02-18T05:24:36.000
dc.date.accessioned 2020-06-30T04:01:13Z
dc.date.available 2020-06-30T04:01:13Z
dc.date.issued 2012-01-01
dc.description.abstract <p>In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.</p>
dc.description.comments <p>This article is from <em>Autophagy </em>8 (2012): 445, doi: <a href="http://dx.doi.org.proxy.lib.iastate.edu/10.4161/auto.19496">10.4161/auto.19496</a>.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/gdcb_las_pubs/125/
dc.identifier.articleid 1129
dc.identifier.contextkey 9839814
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath gdcb_las_pubs/125
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/37790
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/gdcb_las_pubs/125/2012_Bassham_GuidelinesUse.pdf|||Fri Jan 14 19:23:06 UTC 2022
dc.source.uri 10.4161/auto.19496
dc.subject.disciplines Cell Biology
dc.subject.disciplines Developmental Biology
dc.subject.disciplines Genetics and Genomics
dc.subject.keywords autolysosome
dc.subject.keywords autophagosome
dc.subject.keywords flux
dc.subject.keywords LC3
dc.subject.keywords lysosome
dc.subject.keywords phagophore
dc.subject.keywords stress
dc.subject.keywords vacuole
dc.title Guidelines for the use and interpretation of assays for monitoring autophagy
dc.type article
dc.type.genre article
dspace.entity.type Publication
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