Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes

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 us...

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Published in:Autophagy 2008-02, Vol.4 (2), p.151-175
Main Authors: Klionsky, Daniel J., Abeliovich, Hagai, Agostinis, Patrizia, Aliev, Gjumrakch, Ballabio, Andrea, Bamber, Bruce A., Bi, Xiaoning, Biard-Piechaczyk, Martine, Bredesen, Dale E., Cebollero, Eduardo, Cecconi, Francesco, Chu, Charleen T., Chung, Jongkyeong, Clarke, Peter G.H., Clarke, Steven G., Cleveland, John L., Colombo, María Isabel, Coto-Montes, Ana, Deretic, Vojo, Di Sano, Federica, Dinesh-Kumar, Savithramma P., Eissa, N. Tony, DiFiglia, Marian, Dorsey, Frank C., Dröge, Wulf, Dron, Michel, Duszenko, Michael, Esclatine, Audrey, Fésüs, László, Finley, Kim D., Fujisaki, Kozo, Gewirtz, David A., Gohla, Antje, Goldberg, Alfred L., Gonzalez, Ramon, He, You-Wen, Hill, Joseph A., Huang, Wei-Pang, Jäättelä, Marja, Jiang, Xuejun, Jin, Shengkan V., Kang, Chanhee, Kelekar, Ameeta, Kiel, Jan A.K.W., Kimchi, Adi, Kinsella, Timothy J., Kiselyov, Kirill, Kitamoto, Katsuhiko, Komatsu, Masaaki, Kondo, Seiji, Kumar, Rakesh, Laporte, Marianne, Liou, Willisa, Macleod, Kay F., Martinet, Wim, Mautner, Josef, Meléndez, Alicia, Miotto, Giovanni, Mistiaen, Wilhelm P., Münz, Christian, Naqvi, Naweed I., Nishino, Ichizo, Noda, Takeshi, Ogawa, Michinaga, Ozpolat, Bulent, Paglin, Shoshana, Palmer, Glen E., Perlmutter, David H., Piacentini, Mauro, Pinkas-Kramarski, Ronit, Rami, Abdelhaq, Sandri, Marco, Sass, Miklós, Seleverstov, Oleksandr, Settleman, Jeffrey, Shacka, John J., Sibirny, Andrei A., Silva-Zacarin, Elaine C.M., Simonsen, Anne, Smith, Mark A., Srinivas, Vickram, Stenmark, Harald, Sulzer, David, Tabas, Ira, Takeshita, Fumihiko, Tanaka, Keiji, Taylor, J. Paul, Terman, Alexei, Thompson, Craig B., Tolkovsky, Aviva M., Tumanovska, Lesya V., Uchiyama, Yasuo, Ueno, Takashi, Uzcátegui, Néstor L., Vaquero, Eva C., Webster, Paul, Xi, Zhijun, Yue, Zhenyu, Zheng, Xiaoxiang, Deter, Russell L.
Format: Article
Language:English
Subjects:
Animals
Autophagy - physiology
Autophagy-Related Protein 8 Family
Binding
Biology
Bioscience
Calcium
Cancer
Cell
Clinical Laboratory Techniques
Cycle
Data Interpretation, Statistical
Eukaryotic Cells - physiology
Guidelines as Topic
Humans
Landes
Life Sciences
Medicin och hälsovetenskap
Microscopy, Fluorescence - methods
Microtubule-Associated Proteins - metabolism
Models, Biological
Organogenesis
Phagosomes - metabolism
Phagosomes - physiology
Plants - metabolism
Protein Processing, Post-Translational
Protein Transport
Proteins
Saccharomyces cerevisiae Proteins - metabolism
TECHNOLOGY
TEKNIKVETENSKAP
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Summary: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.
ISSN:1554-8627
1554-8635
1554-8635
DOI:10.4161/auto.5338