Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)
Number of Authors: 29172021 (English)In: Autophagy, ISSN 1554-8627, E-ISSN 1554-8635, Vol. 17, no 1Article, review/survey (Refereed) Published
Abstract [en]
In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being 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 autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
Place, publisher, year, edition, pages
Taylor & Francis, 2021. Vol. 17, no 1
Keywords [en]
Autophagosome; cancer; flux; LC3; lysosome; macroautophagy; neurodegeneration; phagophore; stress; vacuole
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:liu:diva-175760DOI: 10.1080/15548627.2020.1797280ISI: 000636121800001PubMedID: 33634751OAI: oai:DiVA.org:liu-175760DiVA, id: diva2:1555638
Note
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Funding: Biotechnology and Biological Sciences Research CouncilUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BBS/E/B/000C0413] Funding Source: Medline; British Heart FoundationBritish Heart Foundation [PG/17/14/32867] Funding Source: Medline; Medical Research CouncilUK Research & Innovation (UKRI)Medical Research Council UK (MRC)European Commission [MR/R009732/1, MR/N022696/1, MR/M00869X/2, MR/S032304/1, MR/S009426/1, G0501003, MR/R025096/1] Funding Source: Medline; NCI NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [R01 CA244144, R01 CA247992, K08 CA230151, R01 CA197398, R01 CA233794, R01 CA190370, R01 CA200310, R01 CA181196] Funding Source: Medline; NEI NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Eye Institute (NEI) [R01 EY027733, R01 EY019643, R01 EY029675, R01 EY026885, P30 EY010572] Funding Source: Medline; NHLBI NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Heart Lung & Blood Institute (NHLBI) [R01 HL085629, R01 HL153614, R01 HL131667, R01 HL141759, R01 HL088256, R01 HL072166, R01 HL118558, R01 HL107594, R01 HL123340, R01 HL154147] Funding Source: Medline; NIAID NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Allergy & Infectious Diseases (NIAID) [R01 AI121244, K08 AI102971, R01 AI136921, R01 AI139046] Funding Source: Medline; NIAMS NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Arthritis & Musculoskeletal & Skin Diseases (NIAMS) [R01 AR077440, R01 AR061370] Funding Source: Medline; NIA NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute on Aging (NIA) [R01 AG062375, P01 AG054407, R01 AG049780, R01 AG062475] Funding Source: Medline; NIDDK NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Diabetes & Digestive & Kidney Diseases (NIDDK) [R01 DK093668, R01 DK108921, R01 DK099558, R01 DK121545, R01 DK112698, R01 DK107220] Funding Source: Medline; NIGMS NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R15 GM102846, R35 GM127029, R35 GM122536, R01 GM115517, R35 GM130331, R35 GM119571, R01 GM118660, R35 GM131919] Funding Source: Medline; NINDS NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Neurological Disorders & Stroke (NINDS) [R01 NS103981, R01 NS115876, P01 NS097197, R25 NS070682, R01 NS091218] Funding Source: Medline; Pancreatic Cancer UK [FLF2015_02_BARTS] Funding Source: Medline; BLRD VA [I01 BX004444, I01 BX004235, I01 BX003803] Funding Source: Medline
2021-05-192021-05-192022-03-23Bibliographically approved