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@ARTICLE{Klionsky:22319,
author = {Klionsky, D.J. and Willbold, D. and (et al.)},
title = {{G}uidelines for the use and interpretation of assays for
monitoring autophagy},
journal = {Autophagy},
volume = {8},
issn = {1554-8627},
address = {Austin, Tex.},
publisher = {Landes Bioscience},
reportid = {PreJuSER-22319},
pages = {445 - 544},
year = {2012},
note = {In a rapidly expanding and highly dynamic field such as
autophagy, it is possible that some authors who should have
been included on this manuscript have been missed. D.J.K.
extends his apologies to researchers in the field of
autophagy who, due to oversight or any other reason, could
not be included. This work was supported by National
Institutes of Health Public Health Service grant GM53396 to
D.J.K. Due to space and other limitations, it is not
possible to include all other sources of financial support.},
abstract = {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.},
keywords = {J (WoSType)},
cin = {ICS-6},
ddc = {570},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {Funktion und Dysfunktion des Nervensystems / BioSoft:
Makromolekulare Systeme und biologische
Informationsverarbeitung},
pid = {G:(DE-Juel1)FUEK409 / G:(DE-Juel1)FUEK505},
shelfmark = {Cell Biology},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000305403400002},
pubmed = {pmid:22966490},
doi = {10.4161/auto.19496},
url = {https://juser.fz-juelich.de/record/22319},
}