Zusammenfassung
Research in autophagy continues to accelerate,(1) and as a result
many new scientists are entering the field. Accordingly, it is important
to establish a standard set of criteria for monitoring macroautophagy
in different organisms. Recent reviews have described the range of
assays that have been used for this purpose.(2,3) There are many
useful and convenient methods that can be used to monitor macroautophagy
in yeast, but relatively few in other model systems, and there is
much confusion regarding acceptable methods to measure macroautophagy
in higher eukaryotes. A key point that needs to be emphasized is
that there is a difference between measurements that monitor the
numbers of autophagosomes versus those that measure flux through
the autophagy pathway; thus, a block in macroautophagy that results
in autophagosome accumulation needs to be differentiated from fully
functional autophagy that includes delivery to, and degradation within,
lysosomes (in most higher eukaryotes) or the vacuole (in plants and
fungi). Here, we present a set of guidelines for the selection and
interpretation of the methods that can be used by investigators who
are attempting to examine macroautophagy and related processes, as
well as by reviewers who need to provide realistic and reasonable
critiques of papers that investigate these processes. This set of
guidelines is 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 verify an
autophagic response.
Nutzer