Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

DANIEL J. KLIONSKY; ET AL.,; ANDREA C. CUMINO; VALERIA A. DÁVILA; JULIA A. LOOS; MARÍA CELESTE NICOLAO

AUTOPHAGY

LANDES BIOSCIENCE

Lugar: Austin, Texas; Año: 2021

1554-8627

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, thistopic has received increasing attention, and many scientists have entered the field. Our knowledge baseand relevant new technologies have also been expanding. Thus, it is important to formulate on a regularbasis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews,there continues to be confusion regarding acceptable methods to evaluate autophagy, especially inmulticellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpretmethods to examine autophagy and related processes, and for reviewers to provide realistic andreasonable critiques of reports that are focused on these processes. These guidelines are not meantto be a dogmatic set of rules, because the appropriateness of any assay largely depends on the questionbeing asked and the system being used. Moreover, no individual assay is perfect for every situation,calling for the use of multiple techniques to properly monitor autophagy in each experimental setting.Finally, several core components of the autophagy machinery have been implicated in distinct autophagicprocesses (canonical and noncanonical autophagy), implying that genetic approaches to blockautophagy should rely on targeting two or more autophagy-related genes that ideally participate indistinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy alsoregulate other cellular pathways including apoptosis, not all of them can be used as a specific marker forbona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy andthe information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technicalinnovation in the field.